Formulations of histone deacetylase inhibitor and uses therof

ABSTRACT

Dosing regimens, methods of treatment, controlled release formulations, and combination therapies that include an HDAC inhibitor, or a pharmaceutically acceptable salt thereof, are described.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.12/761,588 filed Apr. 16, 2010, which claims benefit of U.S. ProvisionalApplication No. 61/170,517 filed on Apr. 17, 2009, both of which areherein incorporated by reference.

FIELD OF THE INVENTION

Pharmaceutical compositions are described that include a histonedeacetylase (HDAC) inhibitor compound for the treatment of cancer.Methods of treatment using the pharmaceutical compositions and dosingregimens are also described.

BACKGROUND OF THE INVENTION

The acetylation state of nucleosomal histones plays an important role inthe regulation of gene expression. Deacetylation of nucleosomal histonesis catalyzed by a group of enzymes known as histone deacetylases(HDACs), of which there are eleven known isoforms. Histone deacetylationleads to chromatin condensation resulting on transcriptional repression,whereas acetylation induces localized relaxation within specificchromosomal regions to allow better access to transcriptional machineryto facilitate transcription.

In tumor cells, selective inhibitors of HDAC enzymes leads to histonehyperacetylation. This alters the transcriptional regulation of a subsetof genes, including many tumor suppressors, genes involved in cell cyclecontrol, cell division and apopotosis. Further, HDAC inhibitors havebeen reported to inhibit tumor growth in vivo. The inhibition of tumorgrowth is accompanied by histone and tubulin hyperacetylation and mayinvolve multiple mechanisms.

HDAC inhibitors block cancer cell proliferation both in vitro and invivo.N-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide(Compound 1) is a hydroxamate-based HDAC inhibitor for use in thetreatment of cancer in a human.

SUMMARY OF THE INVENTION

Pharmaceutical compositions, methods of treating cancer, dosing regimensand combination therapies are disclosed. In one aspect, described is amethod of treating cancer in a human comprising: administering to thehuman a pharmaceutical composition comprising an histone deacetylase(HDAC) inhibitor in cycles consisting of 5 to 9 consecutive days ofdaily administration of the pharmaceutical composition comprising theHDAC inhibitor followed by 2 to 7 consecutive days with noadministration of the pharmaceutical composition comprising the HDACinhibitor. In some embodiments, the method of treating cancer in a humancomprises: administering to the human a pharmaceutical compositioncomprising an histone deacetylase (HDAC) inhibitor in cycles consistingof 5 to 9 consecutive days of daily administration of the pharmaceuticalcomposition comprising the HDAC inhibitor followed by 5 to 7 consecutivedays with no administration of the pharmaceutical composition comprisingthe HDAC inhibitor

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HDAC inhibitorcomprises: daily administration of two immediate release pharmaceuticalcompositions comprising the HDAC inhibitor, wherein the two immediaterelease pharmaceutical compositions are administered consecutively withthe second immediate release pharmaceutical composition beingadministered about 4 to about 6 hours form the first immediate releasepharmaceutical compositions; or daily administration of a singlecontrolled release oral solid dosage pharmaceutical compositioncomprising the HDAC inhibitor.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HDAC inhibitorcomprises daily administration of the HDAC inhibitor in sufficientamount to maintain effective plasma concentrations of the HDAC inhibitorin the human for at least about 6 consecutive hours on the days ofdosing. In some embodiments, the 5 to 9 consecutive days of dailyadministration of the pharmaceutical composition comprising the HDACinhibitor comprises daily administration of the HDAC inhibitor insufficient amount to maintain effective plasma concentrations of theHDAC inhibitor in the human for at least about 6 consecutive hours onthe days of dosing but not exceeding 14 consecutive hours.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HDAC inhibitorcomprises daily administration of the HDAC inhibitor in sufficientamount to maintain effective plasma concentrations of the HDAC inhibitorin the human for about 6 consecutive hours to about 8 consecutive hourson the days of dosing.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HDAC inhibitorcomprises: daily administration of two immediate release pharmaceuticalcompositions comprising the HDAC inhibitor, wherein the two immediaterelease pharmaceutical compositions are administered consecutively 4 to6 hours apart; or daily administration of a single controlled releaseoral solid dosage pharmaceutical composition comprising the HDACinhibitor.

In some embodiments, the single controlled release oral solid dosagepharmaceutical composition comprising the HDAC inhibitor providessubstantially the same in vivo release in the human as two immediaterelease pharmaceutical compositions comprising the HDAC inhibitoradministered consecutively 4 to 6 hours apart.

In some embodiments, the HDAC inhibitor is:N-hydroxy-4-[2-(4-methoxyquinolin-2-ylcarbonylamino)ethoxy]benzamide;N-hydroxy-4-[2S-(trans-cinnamoylamino)butoxy]benzamide;N-hydroxy-4-[2R-(trans-cinnamoylamino)butoxy]benzamide;N-hydroxy-4-{2-[4-(2-methoxyethoxy)quinolin-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-[2S-(benzothiophen-2-ylcarbonylamino)butoxy]benzamide;N-hydroxy-4-{2S-[benzofuran-2-ylcarbonylamino]butoxy}benzamide;N-hydroxy-4-{2-[3-(methoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(i-propoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-{2-[3-(3-hydroxypropoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(2-methoxyethyloxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(pyrrolidin-1-ylmethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(piperidin-1-ylmethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(4-methylpiperazin-1-ylmethyl)benzofuran-2-ylcarbonylamino]-ethoxy}benzamide;N-hydroxy-4-{2-[5-(tetrahydropyran-4-yloxy)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2S-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]butoxy}-benzamide;N-hydroxy-4-{2-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]-1R-methyl-ethoxy}benzamide;N-hydroxy-4-{2-[(3-(benzofuran-2-yl)-4-(dimethylamino)-but-2-enoyl)amino]ethoxy}benzamide;or a pharmaceutically acceptable salt thereof.

In some embodiments, the HDAC inhibitor is the HCl salt ofN-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HCl salt ofN-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamidecomprises: daily administration of two immediate release pharmaceuticalcompositions comprising the HCl salt of Compound 1, wherein the twoimmediate release pharmaceutical compositions are administered 4 to 6hours apart; or daily administration of a single controlled release oralsolid dosage pharmaceutical composition.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HCl salt ofN-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamidecomprises daily administration of about 10 mg to about 300 mg of the HClsalt ofN-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide.

In some embodiments, the cancer is a hematological cancer, solid tumoror a sarcoma. In some embodiments, the cancer is breast cancer, coloncancer, colorectal carcinomas, non-small cell lung cancer, small-celllung cancer, liver cancer, ovarian cancer, prostate cancer, uterinecervix cancer, urinary bladder cancer, gastric carcinoma,gastrointestinal stromal tumor, pancreatic cancer, germ cell tumors,mast cell tumors, neuroblastoma, mastocytosis, testicular cancers,glioblastomas, astrocytomas, B cell lymphoma, T cell lymphoma, Hodgkin'slymphoma, non-Hodgkin's lymphoma, melanoma, myeloma, acute myelocyticleukemia (AML), acute lymphocytic leukemia (ALL), myelodysplasticsyndrome, and chronic myelogenous leukemia.

In some embodiments, the method further comprises administering to thehuman at least one additional therapeutic agent selected fromDNA-damaging agents; topoisomerase I or II inhibitors; alkylatingagents; PARP inhibitors; proteasome inhibitors; RNA/DNA antimetabolites;antimitotics; immunomodulatory agents; antiangiogenics; aromataseinhibitors; hormone-modulating agents; apoptosis inducing agents; kinaseinhibitors; monoclonal antibodies; abarelix; ABT-888; aldesleukin;aldesleukin; alemtuzumab; alitretinoin; allopurinol; altretamine;amifostine anastrozole; arsenic trioxide; asparaginase; azacitidine;AZD-2281; bendamustine; bevacizumab; bexarotene; bleomycin; bortezomib;BSI-201; busulfan; busulfan; calusterone; capecitabine; carboplatin;carfilozib; carmustine; carmustine; celecoxib; cetuximab; chlorambucil;cisplatin; cladribine; clofarabine; cyclophosphamide; cytarabine;cytarabine liposomal; dacarbazine; dactinomycin; darbepoetin alfa;dasatinib; daunorubicin liposomal; daunorubicin; decitabine; denileukin;dexrazoxane; docetaxel; doxorubicin; doxorubicin liposomal;dromostanolone propionate; epirubicin; epoetin alfa; erlotinib;estramustine; etoposide phosphate; etoposide; exemestane; filgrastim;floxuridine; fludarabine; fluorouracil; fulvestrant; gefitinib;gemcitabine; gemtuzumab ozogamicin; goserelin acetate; histrelinacetate; hydroxyurea; Ibritumomab tiuxetan; idarubicin; ifosfamide;imatinib mesylate; interferon alfa 2a; Interferon alfa-2b; irinotecan;lenalidomide; letrozole; leucovorin; leuprolide Acetate; levamisole;lomustine; meclorethamine; megestrol acetate; melphalan; mercaptopurine;methotrexate; methoxsalen; mitomycin C; mitomycin C; mitotane;mitoxantrone; nandrolone phenpropionate; nelarabine; NPI-0052;nofetumomab; oprelvekin; oxaliplatin; paclitaxel; paclitaxelprotein-bound particles; palifermin; pamidronate; panitumumab;pegademase; pegaspargase; pegfilgrastim; pemetrexed disodium;pentostatin; pipobroman; plicamycin, mithramycin; porfimer sodium;procarbazine; quinacrine; RAD001; rasburicase; rituximab; sargramostim;Sargramostim; sorafenib; streptozocin; sunitinib malate; tamoxifen;temozolomide; teniposide; testolactone; thalidomide; thioguanine;thiotepa; topotecan; toremifene; tositumomab; tositumomab/I-131tositumomab; trastuzumab; tretinoin; uracil Mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zoledronate; andzoledronic acid.

In some embodiments, the method further comprises radiation therapy.

In one aspect, described herein is a method of treating cancer in ahuman comprising administering to the human a pharmaceutical compositioncomprising the HCl salt ofN-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide(Compound 1) and at least one pharmaceutically acceptable excipient incycles consisting of 5 to 9 consecutive days of daily administration ofthe pharmaceutical composition comprising the HCl salt of Compound 1followed by 5 to 7 consecutive days with no administration of thepharmaceutical composition comprising the HCl salt of Compound 1. Insome embodiments, the 5 to 9 consecutive days of daily administration ofthe pharmaceutical composition comprising the HCl salt of Compound 1comprises daily administration of two immediate release doses of apharmaceutical composition comprising the HCl salt of Compound 1,wherein the two immediate release doses are administered 4 to 6 hoursapart.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HCl salt of Compound 1comprises administering a single controlled release oral solid dosagepharmaceutical composition as described herein.

In some embodiments, the 5 to 9 consecutive days of daily administrationof the pharmaceutical composition comprising the HCl salt of Compound 1comprises about 10 mg to about 300 mg of the HCl salt of Compound 1.

In one aspect, the method of treating cancer with a HDAC inhibitor asdescribed herein decreases the incidence of grade 4 thrombocytopenia inthe human with cancer.

In one aspect is the use of the HCl salt of Compound 1 in themanufacture of a controlled release pharmaceutical composition for oraladministration to a human with cancer.

In one aspect is the use of a controlled release pharmaceuticalcomposition of the HCl salt of Compound 1 for treating cancer in ahuman.

In one aspect is a dosing regimen of a pharmaceutical composition foruse in the treatment of cancer in a human, wherein the pharmaceuticalcomposition includes Compound 1, or a pharmaceutically acceptable saltthereof and the dosing regimen reduces the incidence of grade 4thrombocytopenia in the human with cancer.

In one aspect, disclosed herein is a controlled release oral soliddosage pharmaceutical composition comprising Compound 1:

or a pharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable excipient, wherein the pharmaceuticalcomposition completely releases Compound 1, or a pharmaceuticallyacceptable salt thereof, over a period of about 6 hours to about 10hours after oral administration to a human.

In one aspect, Compound 1 is present in the pharmaceutical compositionsas the HCl salt.

In some embodiments, the pharmaceutical composition completely releasesCompound 1, HCl: (i) at a constant rate over a period of about 6 hoursto about 10 hours after oral administration to a human; (ii) at adecreasing rate over a period of about 6 hours to about 10 hours afteroral administration to a human; or (iii) in pulses over a period ofabout 6 hours to about 10 hours after oral administration to a human.

In some embodiments, the pharmaceutical composition completely releasesCompound 1, HCl within 10 hours after oral administration to a human.

In some embodiments, the pharmaceutical composition releases less thanabout 10% of Compound 1, HCl in the stomach after oral administration tothe human.

In some embodiments, the pharmaceutical composition does not releaseCompound 1, HCl in the stomach after oral administration to the human.

In some embodiments, the pharmaceutical composition comprises Compound1, HCl in a controlled release matrix.

In some embodiments, the pharmaceutical composition is in the form of atablet with an enteric coating.

In some embodiments, the enteric coating comprises: hydroxypropylcellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose,methyl cellulose, ethyl cellulose, cellulose acetate, cellulose acetatephthalate, cellulose acetate trimellitate, hydroxypropylmethyl cellulosephthalate, cellulose ester-ether phthalate, hydroxypropylcellulosephthalate, alkali salts of cellulose acetate phthalate, alkaline earthsalts of cellulose acetate phthalate, hydroxypropylmethyl cellulosephthalate, cellulose acetate phthalate, carboxymethylcellulose sodium,acrylic acid polymers and copolymers, ethyl acrylate/methylmethacrylate/ethyl trimethylammonium chloride methacrylate terpolymer,methacrylic acid/ethyl acrylate copolymer, methacrylic acid/methylmethacrylate copolymer, polyvinyl pyrrolidone, polyvinyl acetate,polyvinylacetate phthalate, vinylacetate crotonic acid copolymer;shellac, ammoniated shellac, shellac-acetyl alcohol, or shellac n-butylstearate.

In some embodiments, the pharmaceutical composition comprises particlesof Compound 1, HCl.

In some embodiments, the pharmaceutical composition completely releasesCompound 1, HCl in pulses over a period of about 6 hours to about 10hours.

In some embodiments, the pharmaceutical composition comprises at leasttwo different groups of particles of Compound 1, HCl.

In some embodiments, the pharmaceutical composition comprises a firstgroup of delayed release particles of Compound 1, HCl and a secondgroups of delayed release particles of Compound 1, HCl.

In some embodiments, the delayed particles of Compound 1, HCl are in theform of beads, pellets, granules, or minitablets.

In some embodiments, the first group of delayed release particles ofCompound 1, HCl delays the release of Compound 1, HCl by at least 1-2hours after oral administration to a human.

In some embodiments, the second group of delayed release particles ofCompound 1, HCl delays the release of Compound 1, HCl by at least 3-6hours after oral administration to a human.

In some embodiments, the release of Compound 1, HCl from the secondgroup of delayed release particles occurs 2-6 hours following therelease of at least half of the amount of Compound 1, HCl from the firstgroup of delayed release particles after administration to a human.

In some embodiments, the pharmaceutical composition releases Compound 1,HCl in two pulses, where the second pulse of Compound 1, HCl occurs 2 to6 hours after the first pulse of Compound 1, HCl after oraladministration to the human.

In some embodiments, the amount of Compound 1, HCl is the same in thetwo groups of particles.

In some embodiments, the delayed release coating on the first group ofdelayed release particles is different from the delayed release coatingon the second group of delayed release particles.

In some embodiments, the delayed release coatings comprise a pHsensitive coating or a pH insensitive coating.

In some embodiments, the pharmaceutical composition is in the form ofpellets, beads, granules or minitablets in a capsule.

In some embodiments, the pharmaceutical composition is in the form ofpellets, beads, or granules that are compressed into a single tablet.

In some embodiments, the pharmaceutical composition comprises about 10mg to about 300 mg of Compound 1, HCl.

In some embodiments, the pharmaceutical composition provides adose-normalized mean AUC_(0-8 h) from about 0.0035 to about 0.0124(μM·h)/(mg/m²) when orally administered to humans.

In one aspect, the pharmaceutical composition is for use in thetreatment of cancer in a human. In some embodiments, the cancer is ahematological cancer, solid tumor or a sarcoma. In some embodiments, thecancer is breast cancer, colon cancer, colorectal carcinomas, non-smallcell lung cancer, small-cell lung cancer, liver cancer, ovarian cancer,prostate cancer, uterine cervix cancer, urinary bladder cancer, gastriccarcinoma, gastrointestinal stromal tumor, pancreatic cancer, germ celltumors, mast cell tumors, neuroblastoma, mastocytosis, testicularcancers, glioblastomas, astrocytomas, a lymphoma, melanoma, myeloma,acute myelocytic leukemia (AML), acute lymphocytic leukemia (ALL),myelodysplastic syndrome, and chronic myelogenous leukemia. In oneaspect, the cancer is a lymphoma or a leukemia. In one aspect, thecancer is a B cell lymphoma, T cell lymphoma, Hodgkin's lymphoma, ornon-Hodgkin's lymphoma. In one aspect, the cancer is non-Hodgkin'slymphoma.

In some embodiments, the pharmaceutical composition is used incombination with radiation therapy.

In some embodiments, the pharmaceutical composition is used incombination with at least one additional therapeutic agent selected fromDNA-damaging agents; topoisomerase I or II inhibitors; alkylatingagents; PARP inhibitors; proteasome inhibitors; RNA/DNA antimetabolites;antimitotics; immunomodulatory agents; antiangiogenics; aromataseinhibitors; hormone-modulating agents; apoptosis inducing agents; kinaseinhibitors; monoclonal antibodies; abarelix; ABT-888; aldesleukin;aldesleukin; alemtuzumab; alitretinoin; allopurinol; altretamine;amifostine anastrozole; arsenic trioxide; asparaginase; azacitidine;AZD-2281; bendamustine; bevacizumab; bexarotene; bleomycin; bortezomib;BSI-201; busulfan; busulfan; calusterone; capecitabine; carboplatin;carfilozib; carmustine; carmustine; celecoxib; cetuximab; chlorambucil;cisplatin; cladribine; clofarabine; cyclophosphamide; cytarabine;cytarabine liposomal; dacarbazine; dactinomycin; darbepoetin alfa;dasatinib; daunorubicin liposomal; daunorubicin; decitabine; denileukin;dexrazoxane; docetaxel; doxorubicin; doxorubicin liposomal;dromostanolone propionate; epirubicin; epoetin alfa; erlotinib;estramustine; etoposide phosphate; etoposide; exemestane; filgrastim;floxuridine; fludarabine; fluorouracil; fulvestrant; gefitinib;gemcitabine; gemtuzumab ozogamicin; goserelin acetate; histrelinacetate; hydroxyurea; Ibritumomab tiuxetan; idarubicin; ifosfamide;imatinib mesylate; interferon alfa 2a; Interferon alfa-2b; irinotecan;lenalidomide; letrozole; leucovorin; leuprolide Acetate; levamisole;lomustine; meclorethamine; megestrol acetate; melphalan; mercaptopurine;methotrexate; methoxsalen; mitomycin C; mitomycin C; mitotane;mitoxantrone; nandrolone phenpropionate; nelarabine; NPI-0052;nofetumomab; oprelvekin; oxaliplatin; paclitaxel; paclitaxelprotein-bound particles; palifermin; pamidronate; panitumumab;pegademase; pegaspargase; pegfilgrastim; pemetrexed disodium;pentostatin; pipobroman; plicamycin, mithramycin; porfimer sodium;procarbazine; quinacrine; RAD001; rasburicase; rituximab; sargramostim;Sargramostim; sorafenib; streptozocin; sunitinib malate; tamoxifen;temozolomide; teniposide; testolactone; thalidomide; thioguanine;thiotepa; topotecan; toremifene; tositumomab; tositumomab/I-131tositumomab; trastuzumab; tretinoin; uracil Mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zoledronate; andzoledronic acid.

Articles of manufacture, which include packaging material, a HDACinhibitor compound described herein, which is effective for selectivelyinhibiting histone deacetylase activity, within the packaging material,and a label that indicates that the compound or composition, orpharmaceutically acceptable salt, pharmaceutically acceptable N-oxide,pharmaceutically active metabolite, pharmaceutically acceptable prodrug,or pharmaceutically acceptable solvate thereof, is used for inhibitingthe activity of histone deacetylase, or for the treatment, prevention oramelioration of one or more symptoms of a disease or condition thatwould benefit from inhibition of histone deacetylase activity, areprovided.

In some embodiments, pharmaceutical compositions described herein thatinclude a HDAC inhibitor are formulated in a manner that is suitable fororal administration to a human.

In some embodiments, pharmaceutical compositions described herein thatinclude a HDAC inhibitor are formulated in a manner that is suitable forintravenous administration to a human.

Other objects, features and advantages of the methods, compounds, andcompositions described herein will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description and the specific examples, while indicatingspecific embodiments, are given by way of illustration only, sincevarious changes and modifications within the spirit and scope of thedisclosure will become apparent to those skilled in the art from thisdetailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 presents the results of in vitro dose scheduling studies. Theeffect of dose schedule of HDAC inhibitor on tumor cell death ispresented.

FIG. 2 presents the results of modeling dose schedules of Compound 1 andthe effects on Jurkat and HCT116 cell lines.

DETAILED DESCRIPTION

Cancer is considered a disease of genetic defects such as a genemutations and deletions, as well as chromosomal abnormalities, thatresult in the loss of function of tumor suppressor genes and/or gain offunction or hyperactivation of oncogenes.

Cancer is characterized by genome-wide changes in gene expression withinthe tumor. These changes favor a tumor's ability to progress through thecell cycle, to avoid apoptosis, or to become resistant to chemotherapy.HDAC inhibitors are able to reverse several of these changes, andrestore a pattern more like that of a normal cell.

The human genome consists of a complex network of genes which are turnedon or off depending on the needs of the cell. One of the ways in whichgenes are turned on or off is by means of chemical modification ofhistone proteins. Histone proteins are structural components ofchromosomes, and form a scaffold upon which DNA, the genetic material,is arranged. A well studied histone modification is acetylation anddeacetylation, modifications that are catalyzed by a family of enzymesknown as histone acetyl transferases and histone deacetylases.

Inhibition of HDAC enzymes by Compound 1 tips the balance in favor ofthe acetylated state, a state that allows transcription to occur, whichcan be thought of as turning a gene “on”. When a cell is treated withCompound 1, waves of previously silenced genes are initially turned on.Some of these genes are regulators themselves, and will activate orrepress the expression of still other genes. The result is an orchestraof changes to gene expression: some genes being turned on, while othersare kept in the off state.

Following chemotherapy and/or radiation treatment, some patient's tumorsmay turn on certain genes as a strategy by the tumor to adapt to thetherapy and become resistant to cell death. One example of a geneticchange that occurs in many cancers is the activation of the DNA repairgene RAD51. In response to treatment with DNA-damaging chemotherapy orradiation, tumors will often turn on DNA repair genes (including RAD51)as an adaptive strategy to help the tumor repair the DNA damage done bythese agents. In pre-clinical models, Compound 1 was able to turn offRAD51 (and other DNA repair genes), effectively blocking the ability ofthe tumor to repair its damaged DNA, sensitizing the tumor tochemotherapy and radiation.

HDAC Inhibitor Preclinical Activities

In preclinical studies Compound 1 has been found to have anticanceractivities with remarkable tumor specificity. These early studiesprovided important information about the in vitro and in vivo activitiesof Compound 1 and determined the molecular mechanism underlying theanticancer effects.

In Vitro:

Compound 1 is active against many tumor cell lines and is efficacious inmouse models of lung, colon, prostate, pancreatic and brain tumors.

Ex Vivo:

Compound 1 is active in primary human tumors from patients with colon,ovarian, lung and many hematological cancers.

Extensive safety and toxicology studies have been completed in multipleanimal species. The mechanism of action of Compound 1 has been studied,and involves a multi-pronged attack on tumor cells: upregulation of p21and other tumor suppressors and cell cycle genes; induction of reactiveoxygen species and attenuation of anti-oxidant pathways; alterations incalcium homeostasis and increased ER stress; downregulation of DNArepair pathways and increased DNA damage; direct induction of apoptosisvia death receptors and activation of caspases.

Compound 1 is a hydroxamate-based HDAC inhibitor. Described herein arepharmaceutical compositions that include Compound 1, as well aspharmacokinetic and pharmacodynamic strategies, dosing regimens, as wellas methods of treating cancers in a human, either as monotherapy orcombination therapy.

Therapeutic and Pharmacodynamic Effect to Compound 1

In clinical trials involving humans with cancer, Compound 1 in solutionform was administered at 2 mg/kg as a single oral dose and as multiple2-hour IV infusion doses. Systemic exposure measured as AUC_(0-∞) for IVand oral dosing was 5.9 μM*hr and 1.45 μM*hr, respectively, indicatingan oral bioavailability of about 27% in humans.

A therapeutic effect of an HDAC inhibitor (e.g. Compound 1) is achievedin humans with cancer by administering the HDAC inhibitor orally twice aday (with the two doses being administered consecutively about 4 toabout 6 hours apart), orally three time a day (with the doses beingadministered consecutively about 4 to about 6 hours apart),intravenously, or continuously. The aforementioned dosing regimensfacilitate the ability to maintain in the human at least 6 consecutivehours of effective plasma concentrations of the HDAC inhibitor.

A therapeutic effect of Compound 1 is achieved in humans with cancer byadministering Compound 1 (immediate release oral capsules) twice a day,with the two doses being administered about 4 to about 6 hours apart. Insome embodiments, twice a day dosing reduces the incidences ofthrombocytopenia as compared to three times a day dosing.

For therapeutic effect, effective plasma concentrations of Compound 1 inhumans should be maintained for at least 6 consecutive hours, at least 7consecutive hours, or at least 8 consecutive hours each day on days ofdosing. For therapeutic effect, effective plasma concentrations ofCompound 1 in humans should be maintained for at least 6 consecutivehours each day on days of dosing. For therapeutic effect, effectiveplasma concentrations of Compound 1 in humans should be maintained forat least 7 consecutive hours each day on days of dosing. In someembodiments, for therapeutic effect, effective plasma concentrations ofCompound 1 in humans should be maintained for about 6 consecutive hoursto about 8 consecutive hours each day on days of dosing. In someembodiments, effective plasma concentrations of Compound 1 in humans aremaintained for at least 6 consecutive hours but not exceeding 12, 13, or14 consecutive hours on days of dosing. In some embodiments, maintainingthe effective plasma concentrations for at least 6 consecutive hours butnot exceeding 14 consecutive hours of Compound 1 on days of dosingincreases the efficacy of tumor cell growth inhibition and minimizes theincidences of thrombocytopenia. In some embodiments, maintaining theeffective plasma concentrations for about 6 consecutive hours to about 8consecutive hours of Compound 1 on days of dosing increases the efficacyof tumor cell growth inhibition and minimizes the incidences ofthrombocytopenia.

The oral bioavailabilty of Compound 1 in humans, administered asimmediate release capsules or an oral solution, was determined to beabout 27%. A difference in pharmacokinetics was observed in laboratoryanimals between the fasted state the fed state. Compound 1 appears to bepreferentially absorbed in the intestines.

In one aspect, presented herein are methods of providing reliable fortherapeutic and pharmacodynamic effect to Compound 1, or apharmaceutically acceptable salt thereof, that include administeringCompound 1, or a pharmaceutically acceptable salt thereof, in the formof a controlled release formulation. Controlled release formulationsallow for once a day dosing. Controlled release formulations also allowfor the release of the active agent (i.e. Compound 1, or apharmaceutically acceptable salt thereof) in the intestines rather thanin the stomach.

In one aspect, the controlled release formulation is a multi-particulatedrug delivery system. Multi-particulate drug delivery systems are oraldosage forms consisting of a multiplicity of small discrete units, eachexhibiting some desired characteristics. In these systems, the dosage ofthe drug substances is divided on a plurality of subunits, typicallyconsisting of thousands of spherical particles with diameter of0.05-2.00 mm. Multiparticulate dosage forms are pharmaceuticalformulations in which the active substance is present as a number ofsmall independent subunits. To deliver the recommended total dose, thesesubunits are filled into a capsule or compressed into a tablet.Multiparticulates are less dependent on gastric emptying, resulting inless inter and intra-subject variability in gastrointestinal transittime. They are also better distributed and less likely to cause localirritation or be influenced by the presence of food.

Multiparticulate dosage forms offer benefits such as increasedbioavailability, reduced risk of local irritation and predictablegastric emptying. In some embodiments, multiparticulate systems showbetter reproducible pharmacokinetic behavior than conventionalformulations.

After disintegration of a controlled release formulation (e.g. tablet orcapsule) which occurs within a few minutes, the individual subunitparticles pass rapidly through the GI tract. If these subunits havediameters of less than 2 mm, they are able to leave the stomachcontinuously, even if the pylorus is closed. These results in lowerintra and inter individual variability in plasma levels andbioavailability.

Other controlled release oral pharmaceutical dosage forms are able toprovide the same benefits that are observed with a multi-particulatedrug delivery system.

Drug Holiday

Thrombocytopenia is a side effect observed in humans that receivetreatment with a HDAC inhibitor compound. Thrombocytopenia is acondition in which there is a lower-than-normal number of platelets inthe blood. It may result in easy bruising and excessive bleeding fromwounds or bleeding in mucous membranes and other tissues.Thrombocytopenia has typically been reconciled by lowering the dailydose of the HDAC inhibitor compound that is administered to the human.However, a lowering of the daily dose of the HDAC inhibitor compound maynot allow for therapeutic and for therapeutic and pharmacodynamic effectto the HDAC inhibitor compound.

Presented herein are dosing regimens for achieving for therapeutic andpharmacodynamic effect to an HDAC inhibitor with a limited incidence ofGrade 4 thrombocytopenia that include 5-9 consecutive days of dailydosing of an HDAC inhibitor in an amount sufficient to maintaineffective plasma concentrations of the HDAC inhibitor for at least 6consecutive hours on each day of dosing, followed by 5-9 consecutivedays without dosing the HDAC inhibitor. In some embodiments, on the daysof dosing the effective plasma concentrations of the HDAC inhibitor aremaintained for at least 6, at least 7, or at least 8 consecutive hoursbut not exceeding 12, 13, or 14 consecutive hours. In some embodiments,on the days of dosing the HDAC inhibitor is administered in an amountsufficient to maintain effective plasma concentrations of the HDACinhibitor for about 6 consecutive hours to about 8 consecutive hours. Insome embodiments, the HDAC inhibitor is administered orally. In someembodiments, on the days of dosing the effective plasma concentrationsof the HDAC inhibitor are maintained for no more than 12, 13, or 14consecutive hours. In other embodiments, the HDAC inhibitor isadministered parenterally.

Presented herein are dosing regimens for achieving for therapeutic andpharmacodynamic effect to an HDAC inhibitor with a limited incidence ofGrade 4 thrombocytopenia that include: (a) twice a day oral dosing of anHDAC inhibitor (immediate release oral pharmaceutical composition) for 7consecutive days followed by 7 consecutive days without dosing the HDACinhibitor; or (b) once a day dosing of an HDAC inhibitor (controlledrelease oral pharmaceutical composition) for 7 consecutive days followedby 7 consecutive days without dosing the HDAC inhibitor. The foregoingdosing regimen also includes 5-9 consecutive days of dosing with an HDACinhibitor, followed by 2-9 days consecutive days of no dosing with anHDAC inhibitor.

Presented herein are dosing regimens for achieving for therapeutic andpharmacodynamic effect to Compound 1 with a limited incidence of Grade 4thrombocytopenia that include: (a) twice a day oral dosing of Compound 1(immediate release oral pharmaceutical composition) for 7 consecutivedays followed by 7 consecutive days without dosing Compound 1; (b) oncea day dosing of Compound 1 (controlled release oral pharmaceuticalcomposition) for 7 consecutive days followed by 7 consecutive dayswithout dosing Compound 1. The foregoing dosing regimen also includes5-9 consecutive days of dosing with Compound 1, followed by 2-9 daysconsecutive days of no dosing with Compound 1.

The foregoing dosing regimen also includes 5-9 consecutive days ofdosing with Compound 1, followed by 2-9 days consecutive days of nodosing with Compound 1.

HDAC Inhibitor Compounds

N-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide(Compound 1) has the following structure:

In one aspect, Compound 1 is used in the pharmaceutical compositions andmethods disclosed herein as a pharmaceutically acceptable salt. In oneaspect, Compound 1 is used as the hydrochloride salt.

Additional pharmaceutically acceptable salts of Compound 1 include: (a)salts formed when the acidic proton of Compound 1 is replaced by a metalion, such as for example, an alkali metal ion (e.g. lithium, sodium,potassium), an alkaline earth ion (e.g. magnesium, or calcium), or analuminum ion, or is replaced by an ammonium cation (NH₄ ⁺); (b) saltsformed by reacting Compound 1 with a pharmaceutically acceptable organicbase, which includes alkylamines, such as ethanolamine, diethanolamine,triethanolamine, tromethamine, N-methylglucamine, dicyclohexylamine,tris(hydroxymethyl)methylamine, and salts with amino acids such asarginine, lysine, and the like; (c) salts formed by reacting Compound 1with a pharmaceutically acceptable acid, which provides acid additionsalts. Pharmaceutically acceptable acids include hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid,metaphosphoric acid, and the like; or with an organic acid, such as, forexample, acetic acid, propionic acid, hexanoic acid,cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid,malonic acid, succinic acid, malic acid, maleic acid, fumaric acid,trifluoroacetic acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonicacid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, and the like.

Additional pharmaceutically acceptable salts include those described inBerge et al., J. Pharm. Sci. 1977, 66, 1-19; and “Handbook ofPharmaceutical Salts, Properties, and Use,” Stah and Wermuth, Ed.;Wiley-VCH and VHCA, Zurich, 2002.

In some embodiments, sites on the aromatic ring portion of compoundsdescribed herein that are susceptible to various metabolic reactions aremodified such that the various metabolic reactions are reduced,minimized or eliminated. Such modifications include incorporation ofappropriate substituents on the aromatic ring structures, such as, byway of example only, halogens, deuterium, and the like. In one aspect,HDAC inhibitor compounds described herein are deuterated at sitessusceptible to metabolic reactions.

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulae and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the present compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, such as, forexample, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ¹⁸F, ³⁶Cl, respectively.Certain isotopically-labeled compounds described herein, for examplethose into which radioactive isotopes such as ³H and ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Further, substitution with isotopes such as deuterium, i.e., ²H,can afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements.

Other HDAC inhibitor compounds that are contemplated for use in thepharmaceutical compositions, pharmacokinetic strategies, dosingregimens, methods of treatments, and combination therapies include thosecompounds with the structure of Formula (I):

wherein:

-   -   X is —O—, —NR²—, or —S(O)_(n) where n is 0, 1, or 2 and R² is        hydrogen, —CH₃, —CH₂CH₃;    -   Y is ethylene, propylene, 1-methylpropylene, 2-methylpropylene,        —CH(C₂H₅)CH₂—, —CH(CH(CH₃)₂)CH₂—, and —CH(CH₃)CH₂—;    -   R³ is hydrogen, —CH₃, or —CH₂CH₃;    -   Ar is phenyl, naphthyl, quinolinyl, benzofuranyl, benzothienyl,        trans phenylCH═CH— or trans (benzofuran-2-yl)CH═CH—, wherein Ar        is optionally substituted with one or two substituents        independently selected from chloro, fluoro, trifluoromethyl,        methyl, ethyl, methoxy, ethoxy, methylenedioxy, —OH,        1-cyclopropylpiperidin-4-yloxy,        1-(2,2,2-trifluoroethyl)piperidin-4-yloxy,        N,N-dimethylaminomethyl, N,N-diethylaminomethyl,        2-methoxyethoxymethyl, phenoxymethyl, 2-methoxyethoxy,        2-morpholin-4-ylethoxy, pyridin-3-ylmethoxy, 2-hydroxyethoxy,        2-N,N-dimethylaminoethoxy, methoxymethyl, 3-i-propoxymethyl,        morpholin-4-ylmethyl, 3-hydroxypropyloxymethyl,        2-fluorophenoxymethyl, 3-fluorophenoxymethyl,        4-fluorophenoxy-methyl, 3-methoxypropyloxymethyl,        pyridin-4-yloxymethyl, 2,4,6-trifluorophenoxymethyl,        2-oxopyridin-1-ylmethyl, 2,2,2-trifluoroethoxymethyl,        4-imidazol-1-ylphenoxymethyl,        4-[1.2.4-triazin-1-yl-phenoxymethyl, 2-phenylethyl,        pyrrolidin-1-ylmethyl, piperidin-1-ylmethyl,        4-trifluoromethylpiperidin-1-ylmethyl,        4-methylpiperazin-1-ylmethyl, 3,3,3-trifluoropropyloxymethyl,        4-fluorophenylthiomethyl, 4-fluorophenylsulfinylmethyl,        4-fluorophenylsulfonylmethyl, pyridin-3-ylmethyloxymethyl,        tetrahydropyran-4-yloxy, 2,2,2-trifluoroethyloxy,        2-pyrrolidin-1-ylethyloxy, piperidin-4-yloxy,        N-methyl-N-benzylaminomethyl,        N-methyl-N-2-phenylethylaminomethyl, 3-hydroxypropylthiomethyl,        3-hydroxypropylsulfinylmethyl, 3-hydroxypropylsulfonyl-methyl,        N-methyl-N-2-indol-3-ylethylaminomethyl,        2-(4-trifluoromethylphenyl)ethyl,        2-(3-trifluoromethoxyphenyl)ethyl,        N-hydroxyaminocarbonyl-methylaminomethyl, or        3-(2-carboxyethylamino-methyl); or    -   a pharmaceutically acceptable salt thereof.

In some embodiments, Ar is benzofuran-2-yl and is monosubstituted at the3-position of the benzofuran-2-yl ring with N,N-dimethylaminomethyl,N,N-diethylaminomethyl, 2-fluorophenoxymethyl, 3-fluorophenoxymethyl,4-fluorophenoxy-methyl, hydroxyl-4-yloxymethyl,2,4,6-trifluorophenoxy-methyl, 2-oxopyridin-1-ylmethyl,2,2,2-trifluoroethoxy-methyl, 4-imidazol-1-ylphenoxy-methyl,4-[1.2.4-triazin-1-yl-phenoxymethyl, 2-phenylethyl,3-hydroxypropyloxymethyl, 2-methoxyethyloxymethyl,pyrrolidin-1-ylmethyl, piperidin-1-ylmethyl,4-trifluoromethylpiperidin-1-ylmethyl, 4-methylpiperazin-1-ylmethyl,3,3,3-trifluoropropyloxymethyl, 4-fluorophenylthiomethyl,4-fluorophenylsulfinylmethyl, 4-fluorophenylsulfonylmethyl,2-(3-trifluoromethoxyphenylethyl), N-methyl-N-benzylaminomethyl,N-methyl-N-2-phenylethylaminomethyl, 3-hydroxypropyl-thiomethyl,3-hydroxypropylsulfinyl-methyl, 3-hydroxypropylsulfonylmethyl,N-methyl-N-2-indol-3-ylethylaminomethyl,2-(4-trifluoromethylphenyl)ethyl,N-hydroxyaminocarbonyl-methylaminomethyl, or 2-carboxyethylaminomethyl.

In some embodiments, Ar is benzofuran-2-yl and is monosubstituted at the3-position of the benzofuran-2-yl ring with N,N-dimethylaminomethyl,N,N-diethylaminomethyl, 2-methoxyethoxymethyl, methoxymethyl,3-i-propoxymethyl, morpholin-4-ylmethyl, 3-hydroxypropyloxymethyl,3-methoxypropyloxymethyl, pyrrolidin-1-ylmethyl, orpiperidin-1-ylmethyl.

In some embodiments, Ar is benzofuran-2-yl and is substituted at the5-position of the benzofuran-2-yl ring with1-cyclopropylpiperidin-4-yloxy, piperidin-4-yloxy,tetrahydropyran-4-yloxy, 2,2,2-trifluoroethoxy,2-pyrrolidin-1-ylethyloxy, or 1-(2,2,2-trifluoroethyl)piperidin-4-yloxy.

In some embodiments, Ar is trans phenylCH═CH— wherein the phenyl isoptionally substituted with one or two substituents independentlyselected from methyl, ethyl, methoxy, ethoxy, methylenedioxy, or —OH. Insome embodiments, Ar is trans phenylCH═CH—.

In some embodiments, Ar is naphthyl wherein the naphthyl is optionallysubstituted with one or two substituents.

In some embodiments, Ar is quinolinyl wherein the quinolinyl isoptionally substituted with one or two substituents.

In some embodiments, Ar is quinolinyl wherein the quinolinyl isoptionally substituted with one or two substituents independentlyselected from chloro, fluoro, trifluoromethyl, methyl, ethyl, methoxy,ethoxy, methylenedioxy, —OH, 2-methoxyethoxy, 2-hydroxyethoxy,methoxymethyl, 3-i-propoxymethyl, 3-hydroxypropyloxymethyl,3-methoxypropyloxymethyl, or 3,3,3-trifluoropropyloxymethyl.

In some embodiments, X is —O— and R³ is hydrogen.

In some embodiments, X is —S(O)_(n) and R³ is hydrogen.

In some embodiments, Y is ethylene. In some embodiments, Y is ethyleneor —CH(C₂H₅)CH₂—. In some embodiments, Y is —CH(C₂H₅)CH₂—.

In some embodiments, X is —O—; R³ is hydrogen; and Y is ethylene or—CH(C₂H₅)CH₂—.

Yet other HDAC inhibitor compounds that are contemplated for use in thepharmaceutical compositions, pharmacokinetic strategies, dosingregimens, methods of treatments, and combination therapies include thosecompounds with the structure of Formula (II):

wherein:

-   -   X is —O—, —NR²—, or —S(O)_(n) where n is 0, 1, or 2 and R² is        hydrogen, —CH₃, —CH₂CH₃;    -   Y is ethylene, propylene, 1-methylpropylene, 2-methylpropylene,        —CH(C₂H₅)CH₂—, —CH(CH(CH₃)₂)CH₂—, and —CH(CH₃)CH₂—;    -   R³ is hydrogen, —CH₃, or —CH₂CH₃;    -   Ar is phenyl, naphthyl, quinolinyl, benzofuranyl, or        benzothienyl, wherein Ar is optionally substituted with one or        two substituents independently selected from chloro, fluoro,        trifluoromethyl, methyl, ethyl, methoxy, ethoxy, methylenedioxy,        —OH;    -   R⁵ is trifluoromethyl, methyl, ethyl, N,N-dimethylaminomethyl,        N,N-diethylaminomethyl, 2-methoxyethoxymethyl, phenoxymethyl,        methoxymethyl, 3-i-propoxymethyl, morpholin-4-ylmethyl,        3-hydroxypropyloxymethyl, 2-fluorophenoxymethyl,        3-fluorophenoxymethyl, 4-fluorophenoxy-methyl,        3-methoxypropyloxymethyl, pyridin-4-yloxymethyl,        2,4,6-trifluorophenoxymethyl, 2-oxopyridin-1-ylmethyl,        2,2,2-trifluoroethoxymethyl, 4-imidazol-1-ylphenoxymethyl,        2-phenylethyl, pyrrolidin-1-ylmethyl, piperidin-1-ylmethyl,        4-trifluoromethylpiperidin-1-ylmethyl,        4-methylpiperazin-1-ylmethyl, 3,3,3-trifluoropropyloxymethyl,        4-fluorophenylthiomethyl, 4-fluorophenylsulfinylmethyl,        4-fluorophenylsulfonylmethyl, pyridin-3-ylmethyloxymethyl,        N-methyl-N-benzylaminomethyl,        N-methyl-N-2-phenylethylaminomethyl, 3-hydroxypropylthiomethyl,        3-hydroxypropylsulfinylmethyl, 3-hydroxypropylsulfonyl-methyl,        N-methyl-N-2-indol-3-ylethylaminomethyl,        2-(4-trifluoromethylphenyl)ethyl,        2-(3-trifluoromethoxyphenyl)ethyl,        N-hydroxyaminocarbonyl-methylaminomethyl, or        3-(2-carboxyethylamino-methyl); or    -   a pharmaceutically acceptable salt thereof.

In some embodiments, Ar is benzofuranyl.

In some embodiments, R⁵ is N,N-dimethylaminomethyl,N,N-diethylaminomethyl, pyrrolidin-1-ylmethyl, or piperidin-1-ylmethyl.

In some embodiments, the HDAC inhibitor is selected from:N-hydroxy-4-[2-(4-methoxyquinolin-2-ylcarbonylamino)ethoxy]benzamide;N-hydroxy-4-[2S-(trans-cinnamoylamino)butoxy]benzamide;N-hydroxy-4-[2R-(trans-cinnamoylamino)butoxy]benzamide;N-hydroxy-4-{2-[4-(2-methoxyethoxy)quinolin-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-[2S-(benzothiophen-2-ylcarbonylamino)butoxy]-benzamide;N-hydroxy-4-{2S-[benzofuran-2-ylcarbonylamino]butoxy}benzamide;N-hydroxy-4-{2-[3-(methoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-{2-[3-(N,N-dimethylaminomethyl)benzofuran-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-{2-[3-(i-propoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}benzamide;N-hydroxy-4-{2-[3-(3-hydroxypropoxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(2-methoxyethyloxymethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(pyrrolidin-1-ylmethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(piperidin-1-ylmethyl)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[3-(4-methylpiperazin-1-ylmethyl)benzofuran-2-ylcarbonylamino]-ethoxy}benzamide;N-hydroxy-4-{2-[5-(tetrahydropyran-4-yloxy)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]ethoxy}-benzamide;N-hydroxy-4-{2S-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]butoxy}-benzamide;N-hydroxy-4-{2-[5-(2-pyrrolidin-1-ylethyloxy)benzofuran-2-ylcarbonylamino]-1R-methyl-ethoxy}benzamide;andN-hydroxy-4-{2-[(3-(benzofuran-2-yl)-4-(dimethylamino)-but-2-enoyl)amino]-ethoxy}benzamide;or a pharmaceutically acceptable salt thereof.

In some embodiments, the HDAC inhibitor is selected from HDAC inhibitorsdisclosed in WO 2004/092115 or WO 2005/097770, both of which are hereinincorporated by reference.

Forms and Phases

HDAC inhibitors (e.g. Compound 1), including pharmaceutically acceptablesalts thereof, and pharmaceutically acceptable solvates thereof, are invarious forms, including but not limited to, amorphous phase, partiallycrystalline forms, crystalline forms, milled forms, and nano-particulateforms. The crystalline forms are known as polymorphs. Polymorphs includethe different crystal packing arrangements of the same elementalcomposition of a compound. This arrangement can significantly affect thephysiochemical, formulation and processing parameters as well as theshelf life or stability of the substance and excipients. Polymorphsusually have different X-ray diffraction patterns, infrared spectra,melting points, density, hardness, crystal shape, optical and electricalproperties, stability, and solubility. Various factors such as therecrystallization solvent, rate of crystallization, and storagetemperature cause a single crystal form to dominate. In one aspect, acrystalline form of an HDAC inhibitor (e.g. Compound 1) is used in thepharmaceutical compositions disclosed herein. In one aspect, acrystalline form of the HCl salt of Compound 1 is used in thepharmaceutical compositions disclosed herein. In one aspect, amorphousCompound 1 is used in the pharmaceutical compositions disclosed herein.In one aspect, amorphous HCl salt of Compound 1 is used in thepharmaceutical composition disclosed herein.

Terminology

“Bioavailability” refers to the percentage of the weight of an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable salt,dosed that is delivered into the general circulation of the animal orhuman being studied. The total exposure (AUC_((0-∞))) of a drug whenadministered intravenously is usually defined as 100% Bioavailable (F%). “Oral bioavailability” refers to the extent to which an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable salt, isabsorbed into the general circulation when the pharmaceuticalcomposition is taken orally as compared to intravenous injection.

“Blood plasma concentration” refers to the concentration an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable salt, inthe plasma component of blood of a subject. It is understood that theplasma concentration of an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt, may vary significantly betweensubjects, due to variability with respect to metabolism and/orinteractions with other therapeutic agents. In one aspect, the bloodplasma concentration of an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt, varies from subject to subject.Likewise, values such as maximum plasma concentration (C_(max)) or timeto reach maximum plasma concentration (T_(max)), or total area under theplasma concentration time curve (AUC_((0-∞))) vary from subject tosubject. Due to this variability, in one embodiment, the amountnecessary to constitute “a therapeutically effective amount” of an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable salt,varies from subject to subject.

“Effective plasma concentrations” of an HDAC inhibitor refers to amountsof the HDAC inhibitor in the plasma that result in exposure levels thatare effective for treating a cancer.

“Drug absorption” or “absorption” typically refers to the process ofmovement of drug from site of administration of a drug across a barrierinto a blood vessel or the site of action, e.g., a drug moving from thegastrointestinal tract into the portal vein or lymphatic system.

A “measurable serum concentration” or “measurable plasma concentration”describes the blood serum or blood plasma concentration, typicallymeasured in mg, μg, or ng of therapeutic agent per ml, dl, or 1 of bloodserum, absorbed into the bloodstream after administration. As usedherein, measurable plasma concentrations are typically measured in ng/mlor μg/ml.

“Pharmacodynamics” refers to the factors which determine the biologicresponse observed relative to the concentration of drug at a site ofaction.

“Pharmacokinetics” refers to the factors which determine the attainmentand maintenance of the appropriate concentration of drug at a site ofaction.

Pharmaceutical Compositions

In one embodiment, oral pharmaceutical compositions are formulated in aconventional manner using one or more physiologically acceptablecarriers (i.e. inactive ingredients) comprising excipients andauxiliaries which facilitate processing of the active compounds intopreparations which are used pharmaceutically. Suitable techniques,carriers, and excipients include those found within, for example,Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton,Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms andDrug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999),herein incorporated by reference in their entirety.

The term “pharmaceutical composition” refers to a mixture of an activeagent (or ingredient) with other inactive chemical components, such ascarriers, stabilizers, diluents, dispersing agents, suspending agents,thickening agents, coatings and/or excipients. The pharmaceuticalcomposition facilitates administration of the compound to a human. Inone aspect, the active agent is an HDAC inhibitor (e.g. Compound 1). Inone aspect, the active agent is the HCl salt of Compound 1.

“Controlled release” as used herein refers to any release profile thatis not entirely immediate release.

For oral administration, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, such as the HCl salt, areformulated by combining the active compound with pharmaceuticallyacceptable carriers or excipients. Such carriers enable an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof, to be formulated as tablets, powders, pills, capsules, and thelike, for oral ingestion by a patient to be treated.

The pharmaceutical compositions will include at least onepharmaceutically acceptable carrier, diluent or excipient and an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof, as the active ingredient.

The oral solid dosage formulations described herein include particles ofan HDAC inhibitor (e.g. Compound 1), or a pharmaceutically acceptablesalt thereof, existing in crystalline form, amorphous phase,semi-crystalline form, semi-amorphous phase, or mixtures thereof.

In one aspect, pharmaceutical compositions disclosed herein are in theform of an oral solid dosage form. Oral solid dosage forms include:tablets, pills, capsule, powders, mini-tablets, particles, beads,pellets, and the like.

The pharmaceutical compositions described herein include an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof, and one or more of the following: (a) binders; (b) coatings;(c) disintegrants; (d) fillers (diluents); (e) lubricants; (f) glidants(flow enhancers); (g) compression aids; (h) colors; (i) sweeteners; (j)preservatives; (k) suspensing/dispersing agents; (l) filmformers/coatings; (m) flavors; (n) printing inks; (o) gelling agents;(p) a second therapeutically active agent.

In one aspect, pharmaceutical compositions described herein include oneor more of the following in addition to the HDAC inhibitor (e.g.Compound 1), or a pharmaceutically acceptable salt thereof: (a)magnesium stearate; (b) lactose; (c) microcrystalline Cellulose; (d)silicified microcrystalline cellulose; (e) mannitol; (f) starch (corn);(g) silicon dioxide; (h) titanium dioxide; (i) stearic acid; (j) sStarch glycolate; (k) gelatin; (l) talc; (m) sucrose; (n) aspartame; (o)calcium stearate; (p) povidone; (q) pregelatinized starch; (r) hydroxypropyl methylcellulose; (s) OPA products (coatings & inks); (t)croscarmellose; (u) hydroxy propyl cellulose; (v) ethylcellulose; (w)calcium phosphate (dibasic); (x) crospovidone; (y) shellac (and glaze);(z) sodium carbonate.

Also provided herein are pharmaceutical compositions comprising anactive ingredient, or a pharmaceutically acceptable salt, solvate, orprodrug thereof, in a pharmaceutically acceptable vehicle, carrier,diluent, or excipient, or a mixture thereof; and one or more releasecontrolling excipients as described herein. Suitable modified releasedosage vehicles include, but are not limited to, hydrophilic orhydrophobic matrix devices, water-soluble separating layer coatings,enteric coatings, osmotic devices, multi-particulate devices, andcombinations thereof. The pharmaceutical compositions may also comprisenon-release controlling excipients.

Provided herein are pharmaceutical compositions in film-coated dosageforms, which comprise a combination of an active ingredient, or apharmaceutically acceptable salt, solvate, or prodrug thereof; and oneor more tabletting excipients to form a tablet core using conventionaltabletting processes and subsequently coating the core. The tablet corescan be produced using conventional granulation methods, for example wetor dry granulation, with optional comminution of the granules and withsubsequent compression and coating. Granulation methods are described,for example, in Voigt, pages 156-69.

Suitable excipients for the production of granules are, for examplepulverulent fillers optionally having flow-conditioning properties, forexample talcum, silicon dioxide, for example synthetic amorphousanhydrous silicic acid of the Syloid® type (Grace), for example SYLOID244 FP, microcrystalline cellulose, for example of the Avicel® type (FMCCorp.), for example of the types AVICEL PH101, 102, 105, RC581 or RC591, Emcocel® type (Mendell Corp.) or Elcema® type (Degussa);carbohydrates, such as sugars, sugar alcohols, starches or starchderivatives, for example lactose, dextrose, saccharose, glucose,sorbitol, mannitol, xylitol, potato starch, maize starch, rice starch,wheat starch or amylopectin, tricalcium phosphate, calcium hydrogenphosphate or magnesium trisilicate; binders, such as gelatin,tragacanth, agar, alginic acid, cellulose ethers, for examplemethylcellulose, carboxymethylcellulose or hydroxypropylmethylcellulose,polyethylene glycols or ethylene oxide homopolymers, especially having adegree of polymerization of approximately from 2.0×10³ to 1.0×10⁵ and anapproximate molecular weight of about from 1.0×10⁵ to 5.0×10⁶, forexample excipients known by the name Polyox® (Union Carbide),polyvinylpyrrolidone or povidones, especially having a mean molecularweight of approximately 1000 and a degree of polymerization ofapproximately from about 500 to about 2500, and also agar or gelatin;surface-active substances, for example anionic surfactants of the alkylsulfate type, for example sodium, potassium or magnesium n-dodecylsulfate, n-tetradecyl sulfate, n-hexadecyl sulfate or n-octadecylsulfate, of the alkyl ether sulfate type, for example sodium, potassiumor magnesium n-dodecyloxyethyl sulfate, n-tetradecyloxyethyl sulfate,n-hexadecyloxyethyl sulfate or n-octadecyloxyethyl sulfate, or of thealkanesulfonate type, for example sodium, potassium or magnesiumn-dodecanesulfonate, n-tetradecanesulfonate, n-hexadecanesulfonate orn-octadecanesulfonate, or non-ionic surfactants of the fatty acidpolyhydroxy alcohol ester type, such as sorbitan monolaurate,monooleate, monostearate or monopalmitate, sorbitan tristearate ortrioleate, polyoxyethylene adducts of fatty acid polyhydroxy alcoholesters, such as polyoxyethylene sorbitan monolaurate, monooleate,monostearate, monopalmitate, tristearate or trioleate, polyethyleneglycol fatty acid esters, such as polyoxyethyl stearate, polyethyleneglycol 400 stearate, polyethylene glycol 2000 stearate, especiallyethylene oxide/propylene oxide block polymers of the Pluronics® (BWC) orSynperonic® (ICI) type

Further provided herein are pharmaceutical compositions in entericcoated dosage forms, which comprise a combination of an activeingredient, or a pharmaceutically acceptable salt, solvate, or prodrugthereof; and one or more release controlling excipients for use in anenteric coated dosage form. The pharmaceutical compositions may alsocomprise non-release controlling excipients.

Additionally provided are pharmaceutical compositions in a dosage formthat has an instant releasing component and at least one delayedreleasing component, and is capable of giving a discontinuous release ofthe compound in the form of at least two consecutive pulses separated intime from 0.5 hour up to 8 hours. The pharmaceutical compositionscomprise a combination of an active ingredient, and one or more releasecontrolling and non-release controlling excipients, such as thoseexcipients suitable for a disruptable semi-permeable membrane and asswellable substances.

Provided herein also are pharmaceutical compositions in a dosage formfor oral administration to a subject, which comprises a combination ofan active ingredient, or a pharmaceutically acceptable salt, solvate, orprodrug thereof; and one or more pharmaceutically acceptable excipientsor carriers, enclosed in an intermediate reactive layer comprising agastric juice-resistant polymeric layered material partially neutralizedwith alkali and having cation exchange capacity and a gastricjuice-resistant outer layer.

Provided herein are pharmaceutical compositions that comprise an activeingredient, in the form of enteric-coated granules, as delayed-releasecapsules for oral administration.

The pharmaceutical compositions provided herein may be provided inunit-dosage forms or multiple-dosage forms. Unit-dosage forms, as usedherein, refer to physically discrete units suitable for administrationto human and animal subjects and packaged individually as is known inthe art. Each unit-dose contains a predetermined quantity of the activeingredient(s) sufficient to produce the desired therapeutic effect, inassociation with the required pharmaceutical carriers or excipients.Examples of unit-dosage forms include individually packaged tablets andcapsules. Unit-dosage forms may be administered in fractions ormultiples thereof. A multiple-dosage form is a plurality of identicalunit-dosage forms packaged in a single container to be administered insegregated unit-dosage form. Examples of multiple-dosage forms includebottles of tablets or capsules.

Pharmaceutical dosage forms can be formulated in a variety of methodsand can provide a variety of drug release profiles, including immediaterelease, sustained release, and delayed release. In some cases it may bedesirable to prevent drug release after drug administration until acertain amount of time has passed (i.e. timed release), to providesubstantially continuous release over a predetermined time period (i.e.sustained release) or to provide release immediately following drugadministration (i.e., immediate release).

Oral formulations that include an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, are presented in the form of:tablets, capsules, pills, pellets, beads, granules, bulk powders.Capsules include mixtures of the active compound(s) with inert fillersand/or diluents such as the pharmaceutically acceptable starches (e.g.corn, potato or tapioca starch), sugars, artificial sweetening agents,powdered celluloses, such as crystalline and microcrystallinecelluloses, flours, gelatins, gums, etc. Tablet formulations are made byconventional compression, wet granulation or dry granulation methods andutilize pharmaceutically acceptable diluents, binding agents,lubricants, disintegrants, surface modifying agents (includingsurfactants), suspending or stabilizing agents, including, but notlimited to, magnesium stearate, stearic acid, talc, sodium laurylsulfate, microcrystalline cellulose, carboxymethylcellulose calcium,polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, xanthan gum,sodium citrate, complex silicates, calcium carbonate, glycine, dextrin,sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose,kaolin, mannitol, sodium chloride, talc, dry starches and powderedsugar. In some embodiments are surface modifying agents which includenonionic and anionic surface modifying agents. For example, surfacemodifying agents include, but are not limited to, poloxamer 188,benzalkonium chloride, calcium stearate, cetostearyl alcohol,cetomacrogol emulsifying wax, sorbitan esters, colloidal silicondioxide, phosphates, sodium dodecylsulfate, magnesium aluminum silicate,and triethanolamine.

In one aspect, oral formulations described herein utilize standard delayor time release formulations to alter the absorption of the activecompound(s).

Binders or granulators impart cohesiveness to a tablet to ensure thetablet remaining intact after compression. Suitable binders orgranulators include, but are not limited to, starches, such as cornstarch, potato starch, and pre-gelatinized starch (e.g., STARCH 1500);gelatin; sugars, such as sucrose, glucose, dextrose, molasses, andlactose; natural and synthetic gums, such as acacia, alginic acid,alginates, extract of Irish moss, Panwar gum, ghatti gum, mucilage ofisabgol husks, carboxymethylcellulose, methylcellulose,polyvinylpyrrolidone (PVP), Veegum, larch arabogalactan, powderedtragacanth, and guar gum; celluloses, such as ethyl cellulose, celluloseacetate, carboxymethyl cellulose calcium, sodium carboxymethylcellulose, methyl cellulose, hydroxyethylcellulose (HEC),hydroxypropylcellulose (HPC), hydroxypropyl methyl cellulose (HPMC);microcrystalline celluloses, such as AVICEL®-PH-101, AVICEL®-PH-103,AVICEL® RC-581, AVICEL®-PH-105 (FMC Corp., Marcus Hook, Pa.); andmixtures thereof. Suitable fillers include, but are not limited to,talc, calcium carbonate, microcrystalline cellulose, powdered cellulose,dextrates, kaolin, mannitol, silicic acid, sorbitol, starch,pre-gelatinized starch, and mixtures thereof. Binder levels are fromabout 50% to about 99% by weight in the pharmaceutical compositionsprovided herein.

Suitable diluents include, but are not limited to, dicalcium phosphate,calcium sulfate, lactose, sorbitol, sucrose, inositol, cellulose,kaolin, mannitol, sodium chloride, dry starch, and powdered sugar.

Suitable disintegrants include, but are not limited to, agar; bentonite;celluloses, such as methylcellulose and carboxymethylcellulose; woodproducts; natural sponge; cation-exchange resins; alginic acid; gums,such as guar gum and Veegum HV; citrus pulp; cross-linked celluloses,such as croscarmellose; cross-linked polymers, such as crospovidone;cross-linked starches; calcium carbonate; microcrystalline cellulose,such as sodium starch glycolate; polacrilin potassium; starches, such ascorn starch, potato starch, tapioca starch, and pre-gelatinized starch;clays; aligns; and mixtures thereof. The amount of disintegrant in thepharmaceutical compositions provided herein varies upon the type offormulation, and is readily discernible to those of ordinary skill inthe art. In one aspect, the pharmaceutical compositions provided hereininclude from about 0.5 to about 15% or from about 1 to about 5% byweight of a disintegrant.

Suitable lubricants include, but are not limited to, calcium stearate;magnesium stearate; mineral oil; light mineral oil; glycerin; sorbitol;mannitol; glycols, such as glycerol behenate and polyethylene glycol(PEG); stearic acid; sodium lauryl sulfate; talc; hydrogenated vegetableoil, including peanut oil, cottonseed oil, sunflower oil, sesame oil,olive oil, corn oil, and soybean oil; zinc stearate; ethyl oleate; ethyllaureate; agar; starch; lycopodium; silica or silica gels, such asAEROSIL® 200 (W. R. Grace Co., Baltimore, Md.) and CAB-O-SIL® (Cabot Co.of Boston, Mass.); and mixtures thereof. In one aspect, thepharmaceutical compositions provided herein include from about 0.1 toabout 5% by weight of a lubricant.

Suitable glidants include colloidal silicon dioxide, CAB-O-SIL® (CabotCo. of Boston, Mass.), and asbestos-free talc. Coloring agents includeany of the approved, certified, water soluble FD&C dyes, and waterinsoluble FD&C dyes suspended on alumina hydrate, and color lakes andmixtures thereof. A color lake is the combination by adsorption of awater-soluble dye to a hydrous oxide of a heavy metal, resulting in aninsoluble form of the dye.

It should be understood that many carriers and excipients may serveseveral functions, even within the same formulation.

In further embodiments, the pharmaceutical compositions provided hereinmay be provided as compressed tablets, tablet triturates, rapidlydissolving tablets, multiple compressed tablets, or enteric-coatingtablets, sugar-coated, or film-coated tablets.

Enteric-coatings are coatings that resist the action of stomach acid butdissolve or disintegrate in the intestine.

In one aspect, the oral solid dosage form disclosed herein include anenteric coating(s). Enteric coatings include one or more of thefollowing: cellulose acetate phthalate; methyl acrylate-methacrylic acidcopolymers; cellulose acetate succinate; hydroxy propyl methyl cellulosephthalate; hydroxy propyl methyl cellulose acetate succinate(hypromellose acetate succinate); polyvinyl acetate phthalate (PVAP);methyl methacrylate-methacrylic acid copolymers; methacrylic acidcopolymers, cellulose acetate (and its succinate and phthalate version);styrol maleic acid co-polymers; polymethacrylic acid/acrylic acidcopolymer; hydroxyethyl ethyl cellulose phthalate; hydroxypropyl methylcellulose acetate succinate; cellulose acetate tetrahydrophtalate;acrylic resin; shellac.

An enteric coating is a coating put on a tablet, pill, capsule, pellet,bead, granule, particle, etc. so that it doesn't dissolve until itreaches the small intestine.

Sugar-coated tablets are compressed tablets surrounded by a sugarcoating, which may be beneficial in covering up objectionable tastes orodors and in protecting the tablets from oxidation.

Film-coated tablets are compressed tablets that are covered with a thinlayer or film of a water-soluble material. Film coatings include, butare not limited to, hydroxyethylcellulose, sodiumcarboxymethylcellulose, polyethylene glycol 4000, and cellulose acetatephthalate. Film coating imparts the same general characteristics assugar coating. Multiple compressed tablets are compressed tablets madeby more than one compression cycle, including layered tablets, andpress-coated or dry-coated tablets.

The tablet dosage forms may be prepared from the active ingredient inpowdered, crystalline, or granular forms, alone or in combination withone or more carriers or excipients described herein, including binders,disintegrants, controlled-release polymers, lubricants, diluents, and/orcolorants. Flavoring and sweetening agents are especially useful in theformation of chewable tablets and lozenges.

The pharmaceutical compositions provided herein may be provided as softor hard capsules, which can be made from gelatin, methylcellulose,starch, or calcium alginate. The hard gelatin capsule, also known as thedry-filled capsule (DFC), consists of two sections, one slipping overthe other, thus completely enclosing the active ingredient. The softelastic capsule (SEC) is a soft, globular shell, such as a gelatinshell, which is plasticized by the addition of glycerin, sorbitol, or asimilar polyol. The capsules may also be coated as known by those ofskill in the art in order to modify or sustain dissolution of the activeingredient.

Coloring and flavoring agents can be used in all of the above dosageforms.

The pharmaceutical compositions provided herein may be formulated asimmediate or modified release dosage forms, including delayed-,sustained, pulsed-, controlled, targeted-, and programmed-release forms.

Pharmaceutical compositions provided herein are in the form of immediateor modified release dosage forms, including delayed-, sustained,pulsed-, controlled, targeted-, and programmed-release forms.

Controlled Release

In one aspect, the pharmaceutical compositions provided herein are inthe form of a controlled release dosage form. As used herein, the term“controlled release” refers to a dosage form in which the rate or placeof release of the active ingredient(s) is different from that of animmediate dosage form when orally administered. Controlled releasedosage forms include delayed-, extended-, prolonged-, sustained-,pulsatile-, modified-, targeted-, programmed-release. The pharmaceuticalcompositions in controlled release dosage forms are prepared using avariety of modified release devices and methods known to those skilledin the art, including, but not limited to, matrix controlled releasedevices, osmotic controlled release devices, multiparticulate controlledrelease devices, ion-exchange resins, enteric coatings, multilayeredcoatings, and combinations thereof. The release rate of the activeingredient(s) can also be modified by varying the particle sizes.

The pharmaceutical solid oral dosage forms including formulationsdescribed herein, which include an HDAC inhibitor (e.g. Compound 1), ora pharmaceutically acceptable salt thereof, are formulated to provide acontrolled release of the HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof.

In contrast to immediate release compositions, controlled releasecompositions allow delivery of an agent to a human over an extendedperiod of time according to a predetermined profile. Such release ratescan provide therapeutically effective levels of agent for an extendedperiod of time and thereby provide a longer period of pharmacologicresponse. Such longer periods of response provide for many inherentbenefits that are not achieved with the corresponding short acting,immediate release preparations. In one aspect, controlled releasecompositions of an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, provide therapeuticallyeffective levels of the HDAC inhibitor (e.g. Compound 1) for an extendedperiod of time and thereby provide a longer period of pharmacologicresponse.

In some embodiments, the solid dosage forms described herein can beformulated as enteric coated delayed release oral dosage forms, i.e., asan oral dosage form of a pharmaceutical composition as described hereinwhich utilizes an enteric coating to affect release in the smallintestine of the gastrointestinal tract. The enteric coated dosage formis a compressed or molded or extruded tablet/mold (coated or uncoated)containing granules, powder, pellets, beads or particles of the activeingredient and/or other composition components, which are themselvescoated or uncoated. In one aspect, the enteric coated oral dosage formmay is a capsule (coated or uncoated) containing pellets, beads orgranules of the solid carrier or the composition, which are themselvescoated or uncoated.

The term “delayed release” as used herein refers to the delivery so thatthe release can be accomplished at some generally predictable locationin the intestinal tract more distal to that which would have beenaccomplished if there had been no delayed release alterations. In someembodiments the method for delay of release is coating. Any coatingsshould be applied to a sufficient thickness such that the entire coatingdoes not dissolve in the gastrointestinal fluids at pH below about 5,but does dissolve at pH about 5 and above. It is expected that anyanionic polymer exhibiting a pH-dependent solubility profile can be usedas an enteric coating in the practice of the present invention toachieve delivery to the lower gastrointestinal tract. In someembodiments the polymers for use in the present invention are anioniccarboxylic polymers. In other embodiments, the polymers and compatiblemixtures thereof, and some of their properties, include, but are notlimited to:

Shellac, also called purified lac. This coating dissolves in media ofpH>7;

Acrylic polymers. The performance of acrylic polymers (primarily theirsolubility in biological fluids) can vary based on the degree and typeof substitution. Examples of suitable acrylic polymers includemethacrylic acid copolymers and ammonio methacrylate copolymers. TheEudragit series E, L, R, S, RL, RS and NE (Rohm Pharma) are available assolubilized in organic solvent, aqueous dispersion, or dry powders. TheEudragit series RL, NE, and RS are insoluble in the gastrointestinaltract but are permeable and are used primarily for colonic targeting.The Eudragit series E dissolve in the stomach. The Eudragit series L,L-30D and S are insoluble in stomach and dissolve in the intestine;

Cellulose Derivatives. Examples of suitable cellulose derivatives are:ethyl cellulose; reaction mixtures of partial acetate esters ofcellulose with phthalic anhydride. The performance can vary based on thedegree and type of substitution. Cellulose acetate phthalate (CAP)dissolves in pH>6. Aquateric (FMC) is an aqueous based system and is aspray dried CAP psuedolatex with particles <1 μm. Other components inAquateric can include pluronics, Tweens, and acetylated monoglycerides.Other suitable cellulose derivatives include: cellulose acetatetrimellitate (Eastman); methylcellulose (Pharmacoat, Methocel);hydroxypropylmethyl cellulose phthalate (HPMCP); hydroxypropylmethylcellulose succinate (HPMCS); and hydroxypropylmethylcellulose acetatesuccinate (e.g., AQOAT (Shin Etsu)). The performance can vary based onthe degree and type of substitution. For example, HPMCP such as, HP-50,HP-55, HP-555, HP-55F grades are suitable. The performance can varybased on the degree and type of substitution. For example, suitablegrades of hydroxypropylmethylcellulose acetate succinate include, butare not limited to, AS-LG (LF), which dissolves at pH 5, AS-MG (MF),which dissolves at pH 5.5, and AS-HG (HF), which dissolves at higher pH.These polymers are offered as granules, or as fine powders for aqueousdispersions;

Poly Vinyl Acetate Phthalate (PVAP). PVAP dissolves in pH>5, and it ismuch less permeable to water vapor and gastric fluids.

In some embodiments, the coating can, and usually does, contain aplasticizer and possibly other coating excipients such as colorants,talc, and/or magnesium stearate, which are well known in the art.Suitable plasticizers include triethyl citrate (Citroflex 2), triacetin(glyceryl triacetate), acetyl triethyl citrate (Citroflec A2), Carbowax400 (polyethylene glycol 400), diethyl phthalate, tributyl citrate,acetylated monoglycerides, glycerol, fatty acid esters, propyleneglycol, and dibutyl phthalate. In particular, anionic carboxylic acrylicpolymers usually will contain 10-25% by weight of a plasticizer,especially dibutyl phthalate, polyethylene glycol, triethyl citrate andtriacetin. Conventional coating techniques such as spray or pan coatingare employed to apply coatings. The coating thickness must be sufficientto ensure that the oral dosage form remains intact until the desiredsite of topical delivery in the intestinal tract is reached.

Colorants, detackifiers, surfactants, antifoaming agents, lubricants(e.g., carnuba wax or PEG) may be added to the coatings besidesplasticizers to solubilize or disperse the coating material, and toimprove coating performance and the coated product.

A particularly suitable methacrylic copolymer is Eudragit L®,particularly L-30D® and Eudragit 100-55®, manufactured by Rohm Pharma,Germany. In Eudragit L-30D®, the ratio of free carboxyl groups to estergroups is approximately 1:1. Further, the copolymer is known to beinsoluble in gastrointestinal fluids having pH below 5.5, generally1.5-5.5, i.e., the pH generally present in the fluid of the uppergastrointestinal tract, but readily soluble or partially soluble at pHabove 5.5, i.e., the pH values present in the small intestine.

In some embodiments, materials include shellac, acrylic polymers,cellulosic derivatives, polyvinyl acetate phthalate, and mixturesthereof. In other embodiments, materials include Eudragit® series E, L,RL, RS, NE, L, L300, S, 100-55, cellulose acetate phthalate, Aquateric,cellulose acetate trimellitate, ethyl cellulose, hydroxypropyl methylcellulose phthalate, hydroxypropyl methyl cellulose acetate succinate,poly vinyl acetate phthalate, and Cotteric.

For some types of drugs, it is preferred to release the drug in“pulses,” wherein a single dosage form provides for an initial dose ofdrug followed by a release-free interval, after which a second dose ofdrug is released, followed by one or more additional release-freeintervals and drug release “pulses.” Alternatively, no drug is releasedfor a period of time after administration of the dosage form, afterwhich a dose of drug is released, followed by one or more additionalrelease-free intervals and drug release “pulses.”

Pulsatile drug delivery is useful, for example, with active agents thathave short half-lives are administered two or three times daily, withactive agents that are extensively metabolized presystemically, and withactive agents that should maintain certain plasma levels in order haveoptimized pharmacodynamic effects.

A pulsatile dosage form is capable of providing one or more immediaterelease pulses at predetermined time points after a controlled lag timeor at specific sites. Pulsatile dosage forms including the formulationsdescribed herein, which include an HDAC inhibitor (e.g. Compound 1), ora pharmaceutically acceptable salt thereof, is administered using avariety of pulsatile formulations that have been described. For example,such formulations include, but are not limited to, those described inU.S. Pat. Nos. 5,011,692, 5,017,381, 5,229,135, 5,840,329, 4,871,549,5,260,068, 5,260,069, 5,508,040, 5,567,441 and 5,837,284. In oneembodiment, the controlled release dosage form is pulsatile releasesolid oral dosage form including at least two groups of particles, (i.e.multiparticulate) each containing the formulation described herein. Thefirst group of particles provides a substantially immediate dose of anHDAC inhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof, upon ingestion by a mammal. The first group of particles can beeither uncoated or include a coating and/or sealant. The second group ofparticles includes coated particles, which includes from about 2% toabout 75%, preferably from about 2.5% to about 70%, and more preferablyfrom about 40% to about 70%, by weight of the total dose of an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof, in said formulation, in admixture with one or more binders. Thecoating includes a pharmaceutically acceptable ingredient in an amountsufficient to provide a delay of from about 2 hours to about 7 hoursfollowing ingestion before release of the second dose. Suitable coatingsinclude one or more differentially degradable coatings such as, by wayof example only, pH sensitive coatings (enteric coatings) such asacrylic resins (e.g., Eudragit® EPO, Eudragit® L30D-55, Eudragit® FS 30DEudragit® L100-55, Eudragit® L100, Eudragit® 5100, Eudragit® RD100,Eudragit® E100, Eudragit® L12.5, Eudragit® S 12.5, and Eudragit® NE30D,Eudragit® NE 40D) either alone or blended with cellulose derivatives,e.g., ethylcellulose, or non-enteric coatings having variable thicknessto provide differential release of the formulation that includes an HDACinhibitor (e.g. Compound 1), or a pharmaceutically acceptable saltthereof.

Multiparticulate Controlled Release Devices

In some embodiments, the pharmaceutical compositions described hereinare multiparticulate controlled release devices, which include amultiplicity of particles, granules, or pellets, ranging from about 10μm to about 3 mm, about 50 μm to about 2.5 mm, or from about 100 μm toabout 1 mm in diameter. Such multiparticulates are made bywet-granulation, dry-granulation, extrusion/spheronization,roller-compaction, melt-congealing, by spray-coating seed cores, andcombinations thereof. See, for example, Multiparticulate Oral DrugDelivery; Marcel Dekker: 1994; and Pharmaceutical PelletizationTechnology; Marcel Dekker: 1989.

Other excipients or carriers as described herein are blended with thepharmaceutical compositions to aid in processing and forming themultiparticulates. The resulting particles may themselves constitute themultiparticulate device or may be coated by various film-formingmaterials, such as enteric polymers, water-swellable, and water-solublepolymers. The multiparticulates can be further processed as a capsule ora tablet.

Intestinal protective drug absorption system (IPDAS) is amultiparticulate tablet technology that consists of high densitycontrolled release beads that are compressed into a tablet form. Thebeads may be manufactured by techniques such as extrusion spheronizationand controlled release can be achieved with the use of different polymersystems to coat the resultant beads. Alternatively, the drug can also becoated onto an inert carrier such as non-pareil seeds to produce instantrelease multiparticulates. Controlled release can be achieved by theformation of a polymeric membrane onto these instant releasemultparticulates. Once an IPDAS tablet is ingested, it rapidlydisintegrates and disperses beads containing the drug in the stomachwhich subsequently pass into the duodenum and along the gastrointestinaltract in a controlled and gradual manner, independent of the feedingstate. Release of active ingredient from the multiparticulates occursthrough a process of diffusion either through the polymeric membraneand/or the micro matrix of the polymer/active ingredient formed in theextruded/spheronized multiparticulates. The intestinal protection ofIPDAS is by virtue of the multiparticulate nature of the formulationwhich ensures wide dispersion of drug throughout the gastrointestinaltract.

Spheroidal oral drug absorption system (SODAS) is a multiparticulatetechnology that enables the production of customized dosage forms andresponds directly to individual drug candidate needs. It can provide anumber of tailored drugs release profiles including immediate release ofdrug followed by sustained release to give rise to a fast onset ofaction which is maintained for at least 12 hours. Alternatively, theopposite scenario can be achieved where drug release is delayed for anumber of hours.

Programmable oral drug absorption system (PRODAS) is presented as anumber of mini tablets contained in hard gelatin capsule. It thuscombines the benefits of tableting technology within a capsule. It ispossible to incorporate many different minitablets, each one formulatedindividually and programmed to release drug at different sites withinthe gastrointestinal tract. These combinations may include immediaterelease, delayed release, and/or controlled release mini tablets. It isalso possible to incorporate mini tablets of different sizes so thathigh drug loading is possible. Their size ranges usually from 1.5-4 mmin diameter.

Many other types of controlled release systems known to those ofordinary skill in the art and are suitable for use with the formulationsdescribed herein. Examples of such delivery systems include, e.g.,polymer-based systems, such as polylactic and polyglycolic acid,plyanhydrides and polycaprolactone; porous matrices, nonpolymer-basedsystems that are lipids, including sterols, such as cholesterol,cholesterol esters and fatty acids, or neutral fats, such as mono-, di-and triglycerides; hydrogel release systems; silastic systems;peptide-based systems; wax coatings, bioerodible dosage forms,compressed tablets using conventional binders and the like. See, e.g.,Liberman et al., Pharmaceutical Dosage Forms, 2 Ed., Vol. 1, pp. 209-214(1990); Singh et al., Encyclopedia of Pharmaceutical Technology, 2^(nd)Ed., pp. 751-753 (2002); U.S. Pat. Nos. 4,327,725, 4,624,848, 4,968,509,5,461,140, 5,456,923, 5,516,527, 5,622,721, 5,686,105, 5,700,410,5,977,175, 6,465,014 and 6,932,983.

Matrix Controlled Release Devices

In some embodiments, the pharmaceutical compositions provided herein isin a modified release dosage form that is fabricated using a matrixcontrolled release device known to those skilled in the art (see, Takadaet al in “Encyclopedia of Controlled Drug Delivery,” Vol. 2, Mathiowitzed., Wiley, 1999).

In one embodiment, the pharmaceutical compositions provided herein in amodified release dosage form is formulated using an erodible matrixdevice, which is water-swellable, erodible, or soluble polymers,including synthetic polymers, and naturally occurring polymers andderivatives, such as polysaccharides and proteins.

Materials useful in forming an erodible matrix include, but are notlimited to, chitin, chitosan, dextran, and pullulan; gum agar, gumarabic, gum karaya, locust bean gum, gum tragacanth, carrageenans, gumghatti, guar gum, xanthan gum, and scleroglucan; starches, such asdextrin and maltodextrin; hydrophilic colloids, such as pectin;phosphatides, such as lecithin; alginates; propylene glycol alginate;gelatin; collagen; and cellulosics, such as ethyl cellulose (EC),methylethyl cellulose (MEC), carboxymethyl cellulose (CMC), CMEC,hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), celluloseacetate (CA), cellulose propionate (CP), cellulose butyrate (CB),cellulose acetate butyrate (CAB), CAP, CAT, hydroxypropyl methylcellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl methyl cellulose acetatetrimellitate (HPMCAT), and ethylhydroxy ethylcellulose (EHEC); polyvinylpyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerol fatty acidesters; polyacrylamide; polyacrylic acid; copolymers of ethacrylic acidor methacrylic acid (EUDRAGIT®, Rohm America, Inc., Piscataway, N.J.);poly(2-hydroxyethyl-methacrylate); polylactides; copolymers ofL-glutamic acid and ethyl-L-glutamate; degradable lactic acid-glycolicacid copolymers; poly-D-(−)-3-hydroxybutyric acid; and other acrylicacid derivatives, such as homopolymers and copolymers ofbutylmethacrylate, methylmethacrylate, ethylmethacrylate, ethylacrylate,(2-dimethylaminoethyl)methacrylate, and(trimethylaminoethyl)methacrylate chloride.

In some embodiments, the pharmaceutical compositions are formulated witha non-erodible matrix device. The active ingredient(s) is dissolved ordispersed in an inert matrix and is released primarily by diffusionthrough the inert matrix once administered. Materials suitable for useas a non-erodible matrix device included, but are not limited to,insoluble plastics, such as polyethylene, polypropylene, polyisoprene,polyisobutylene, polybutadiene, polymethylmethacrylate,polybutylmethacrylate, chlorinated polyethylene, polyvinylchloride,methyl acrylate-methyl methacrylate copolymers, ethylene-vinylacetatecopolymers, ethylene/propylene copolymers, ethylene/ethyl acrylatecopolymers, vinylchloride copolymers with vinyl acetate, vinylidenechloride, ethylene and propylene, ionomer polyethylene terephthalate,butyl rubber epichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer, polyvinyl chloride, plasticizednylon, plasticized polyethyleneterephthalate, natural rubber, siliconerubbers, polydimethylsiloxanes, silicone carbonate copolymers, and;hydrophilic polymers, such as ethyl cellulose, cellulose acetate,crospovidone, and cross-linked partially hydrolyzed polyvinyl acetate,and fatty compounds, such as carnauba wax, microcrystalline wax, andtriglycerides.

In a matrix controlled release system, the desired release kinetics canbe controlled, for example, via the polymer type employed, the polymerviscosity, the particle sizes of the polymer and/or the activeingredient(s), the ratio of the active ingredient(s) versus the polymer,and other excipients or carriers in the compositions.

In one aspect, modified release dosage forms are prepared by methodsknown to those skilled in the art, including direct compression, dry orwet granulation followed by compression, melt-granulation followed bycompression.

In some embodiments, a matrix controlled release system includes anenteric coating so that no drug is released in the stomach.

Osmotic Controlled Release Devices

In some embodiments, the pharmaceutical compositions provided herein ina modified release dosage form is fabricated using an osmotic controlledrelease device, including one-chamber system, two-chamber system,asymmetric membrane technology (AMT), and extruding core system (ECS).In general, such devices have at least two components: (a) the corewhich contains the active ingredient(s); and (b) a semipermeablemembrane with at least one delivery port, which encapsulates the core.The semipermeable membrane controls the influx of water to the core froman aqueous environment of use so as to cause drug release by extrusionthrough the delivery port(s).

In addition to the active ingredient(s), the core of the osmotic deviceoptionally includes an osmotic agent, which creates a driving force fortransport of water from the environment of use into the core of thedevice. One class of osmotic agents water-swellable hydrophilicpolymers, which are also referred to as “osmopolymers” and “hydrogels,”including, but not limited to, hydrophilic vinyl and acrylic polymers,polysaccharides such as calcium alginate, polyethylene oxide (PEO),polyethylene glycol (PEG), polypropylene glycol (PPG),poly(2-hydroxyethyl methacrylate), poly(acrylic) acid, poly(methacrylic)acid, polyvinylpyrrolidone (PVP), crosslinked PVP, polyvinyl alcohol(PVA), PVA/PVP copolymers, PVA/PVP copolymers with hydrophobic monomerssuch as methyl methacrylate and vinyl acetate, hydrophilic polyurethanescontaining large PEO blocks, sodium croscarmellose, carrageenan,hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC),hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC) andcarboxyethyl, cellulose (CEC), sodium alginate, polycarbophil, gelatin,xanthan gum, and sodium starch glycolate.

The other class of osmotic agents are osmogens, which are capable ofimbibing water to affect an osmotic pressure gradient across the barrierof the surrounding coating. Suitable osmogens include, but are notlimited to, inorganic salts, such as magnesium sulfate, magnesiumchloride, calcium chloride, sodium chloride, lithium chloride, potassiumsulfate, potassium phosphates, sodium carbonate, sodium sulfite, lithiumsulfate, potassium chloride, and sodium sulfate; sugars, such asdextrose, fructose, glucose, inositol, lactose, maltose, mannitol,raffinose, sorbitol, sucrose, trehalose, and xylitol; organic acids,such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleicacid, sebacic acid, sorbic acid, adipic acid, edetic acid, glutamicacid, p-toluenesulfonic acid, succinic acid, and tartaric acid; urea;and mixtures thereof.

Osmotic agents of different dissolution rates may be employed toinfluence how rapidly the active ingredient(s) is initially deliveredfrom the dosage form. For example, amorphous sugars, such as MannogemeEZ (SPI Pharma, Lewes, Del.) can be used to provide faster deliveryduring the first couple of hours to promptly produce the desiredtherapeutic effect, and gradually and continually release of theremaining amount to maintain the desired level of therapeutic orprophylactic effect over an extended period of time. In this case, theactive ingredient(s) is released at such a rate to replace the amount ofthe active ingredient metabolized and excreted.

The core may also include a wide variety of other excipients andcarriers as described herein to enhance the performance of the dosageform or to promote stability or processing.

Materials useful in forming the semi-permeable membrane include variousgrades of acrylics, vinyls, ethers, polyamides, polyesters, andcellulosic derivatives that are water-permeable and water-insoluble atphysiologically relevant pHs, or are susceptible to being renderedwater-insoluble by chemical alteration, such as crosslinking. Examplesof suitable polymers useful in forming the coating, include plasticized,unplasticized, and reinforced cellulose acetate (CA), cellulosediacetate, cellulose triacetate, CA propionate, cellulose nitrate,cellulose acetate butyrate (CAB), CA ethyl carbamate, CAP, CA methylcarbamate, CA succinate, cellulose acetate trimellitate (CAT), CAdimethylaminoacetate, CA ethyl carbonate, CA chloroacetate, CA ethyloxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-toluenesulfonate, agar acetate, amylose triacetate, beta glucan acetate, betaglucan triacetate, acetaldehyde dimethyl acetate, triacetate of locustbean gum, hydroxylated ethylene-vinylacetate, EC, PEG, PPG, PEG/PPGcopolymers, PVP, HEC, HPC, CMC, CMEC, HPMC, HPMCP, HPMCAS, HPMCAT,poly(acrylic) acids and esters and poly-(methacrylic) acids and estersand copolymers thereof, starch, dextran, dextrin, chitosan, collagen,gelatin, polyalkenes, polyethers, polysulfones, polyethersulfones,polystyrenes, polyvinyl halides, polyvinyl esters and ethers, naturalwaxes, and synthetic waxes.

Semi-permeable membrane may also be a hydrophobic microporous membrane,wherein the pores are substantially filled with a gas and are not wettedby the aqueous medium but are permeable to water vapor, as disclosed inU.S. Pat. No. 5,798,119. Such hydrophobic but water-vapor permeablemembrane are typically composed of hydrophobic polymers such aspolyalkenes, polyethylene, polypropylene, polytetrafluoroethylene,polyacrylic acid derivatives, polyethers, polysulfones,polyethersulfones, polystyrenes, polyvinyl halides, polyvinylidenefluoride, polyvinyl esters and ethers, natural waxes, and syntheticwaxes.

The delivery port(s) on the semi-permeable membrane may be formedpost-coating by mechanical or laser drilling. Delivery port(s) may alsobe formed in situ by erosion of a plug of water-soluble material or byrupture of a thinner portion of the membrane over an indentation in thecore. In addition, delivery ports may be formed during coating process,as in the case of asymmetric membrane coatings of the type disclosed inU.S. Pat. Nos. 5,612,059 and 5,698,220.

The total amount of the active ingredient(s) released and the releaserate can substantially by modulated via the thickness and porosity ofthe semi-permeable membrane, the composition of the core, and thenumber, size, and position of the delivery ports.

The pharmaceutical compositions in an osmotic controlled-release dosageform may further comprise additional conventional excipients or carriersas described herein to promote performance or processing of theformulation.

The osmotic controlled-release dosage forms can be prepared according toconventional methods and techniques known to those skilled in the art(see, Remington: The Science and Practice of Pharmacy, supra; Santus andBaker, J. Controlled Release 1995, 35, 1-21; Verma et al., DrugDevelopment and Industrial Pharmacy 2000, 26, 695-708; Verma et al., J.Controlled Release 2002, 79, 7-27).

In other embodimthe pharmaceutical compositions provided herein areformulated as AMT controlled-release dosage form, which comprises anasymmetric osmotic membrane that coats a core comprising the activeingredient(s) and other pharmaceutically acceptable excipients orcarriers. See U.S. Pat. No. 5,612,059 and WO 2002/17918. The AMTcontrolled-release dosage forms can be prepared according toconventional methods and techniques known to those skilled in the art,including direct compression, dry granulation, wet granulation, and adip-coating method.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated as ESC controlled-release dosage form, which comprises anosmotic membrane that coats a core comprising the active ingredient(s),a hydroxylethyl cellulose, and other pharmaceutically acceptableexcipients or carriers.

Multilayered Tablets

In one aspect, the controlled release formulation is in the form of amultilayered tablet. Multilayered tablets include an inert core, ontowhich is applied a layered of drug (plus optional excipients), followedby an enteric coating. A second layer of drug is applied onto the firstenteric coating followed by a second enteric coating on the second layerof drug. The enteric coatings should ensure that the release of drugfrom each layer is separated in time by at least 3-6 hours.

Immediate Release

In some embodiments, the pharmaceutical compositions provided herein inan immediate release dosage form are capable of releasing not less than75% of the therapeutically active ingredient or combination and/or meetthe disintegration or dissolution requirements for immediate releasetablets of the particular therapeutic agents or combination included inthe tablet core, as set forth in USP XXII, 1990 (The United StatesPharmacopeia.). Immediate release pharmaceutical compositions includecapsules, tablets, oral solutions, powders, beads, pellets, particles,and the like.

Parenteral Administration

In some embodiments, the pharmaceutical compositions provided herein maybe administered parenterally by injection, infusion, or implantation,for local or systemic administration. Parenteral administration, as usedherein, include intravenous, intraarterial, intraperitoneal,intrathecal, intraventricular, intraurethral, intrasternal,intracranial, intramuscular, intrasynovial, and subcutaneousadministration.

In other embodiments, the pharmaceutical compositions provided hereinmay be formulated in any dosage forms that are suitable for parenteraladministration, including solutions, suspensions, emulsions, micelles,liposomes, microspheres, nanosystems, and solid forms suitable forsolutions or suspensions in liquid prior to injection. Such dosage formscan be prepared according to conventional methods known to those skilledin the art of pharmaceutical science (see, Remington: The Science andPractice of Pharmacy, supra).

The pharmaceutical compositions intended for parenteral administrationmay include one or more pharmaceutically acceptable carriers andexcipients, including, but not limited to, aqueous vehicles,water-miscible vehicles, non-aqueous vehicles, antimicrobial agents orpreservatives against the growth of microorganisms, stabilizers,solubility enhancers, isotonic agents, buffering agents, antioxidants,local anesthetics, suspending and dispersing agents, wetting oremulsifying agents, complexing agents, sequestering or chelating agents,cryoprotectants, lyoprotectants, thickening agents, pH adjusting agents,and inert gases.

Suitable aqueous vehicles include, but are not limited to, water,saline, physiological saline or phosphate buffered saline (PBS), sodiumchloride injection, Ringers injection, isotonic dextrose injection,sterile water injection, dextrose and lactated Ringers injection.Non-aqueous vehicles include, but are not limited to, fixed oils ofvegetable origin, castor oil, corn oil, cottonseed oil, olive oil,peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil,hydrogenated vegetable oils, hydrogenated soybean oil, and medium-chaintriglycerides of coconut oil, and palm seed oil. Water-miscible vehiclesinclude, but are not limited to, ethanol, 1,3-butanediol, liquidpolyethylene glycol (e.g., polyethylene glycol 300 and polyethyleneglycol 400), propylene glycol, glycerin, N-methyl-2-pyrrolidone,dimethylacetamide, and dimethylsulfoxide.

Suitable antimicrobial agents or preservatives include, but are notlimited to, phenols, cresols, mercurials, benzyl alcohol, chlorobutanol,methyl and propyl p-hydroxybenzates, thimerosal, benzalkonium chloride,benzethonium chloride, methyl- and propyl-parabens, and sorbic acid.Suitable isotonic agents include, but are not limited to, sodiumchloride, glycerin, and dextrose. Suitable buffering agents include, butare not limited to, phosphate and citrate. Suitable antioxidants arethose as described herein, including bisulfite and sodium metabisulfite.Suitable local anesthetics include, but are not limited to, procainehydrochloride. Suitable suspending and dispersing agents are those asdescribed herein, including sodium carboxymethylcelluose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Suitable emulsifying agentsinclude those described herein, including polyoxyethylene sorbitanmonolaurate, polyoxyethylene sorbitan monooleate 80, and triethanolamineoleate. Suitable sequestering or chelating agents include, but are notlimited to EDTA. Suitable pH adjusting agents include, but are notlimited to, sodium hydroxide, hydrochloric acid, citric acid, and lacticacid. Suitable complexing agents include, but are not limited to,cyclodextrins, including α-cyclodextrin, β-cyclodextrin,hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin, andsulfobutylether 7-β-cyclodextrin (CAPTISOL®, CyDex, Lenexa, Kans.).

In some embodiments, the pharmaceutical compositions provided herein maybe formulated for single or multiple dosage administration. The singledosage formulations are packaged in an ampule, a vial, or a syringe. Themultiple dosage parenteral formulations must contain an antimicrobialagent at bacteriostatic or fungistatic concentrations. All parenteralformulations must be sterile, as known and practiced in the art.

In one embodiment, the pharmaceutical compositions are provided asready-to-use sterile solutions. In another embodiment, thepharmaceutical compositions are provided as sterile dry solubleproducts, including lyophilized powders and hypodermic tablets, to bereconstituted with a vehicle prior to use. In yet another embodiment,the pharmaceutical compositions are provided as ready-to-use sterilesuspensions. In yet another embodiment, the pharmaceutical compositionsare provided as sterile dry insoluble products to be reconstituted witha vehicle prior to use. In still another embodiment, the pharmaceuticalcompositions are provided as ready-to-use sterile emulsions.

Pharmacokinetic Analysis

In one embodiment, any standard pharmacokinetic protocol is used todetermine blood plasma concentration profile in humans followingadministration of a formulation described herein that includes an HDACinhibitor (e.g. Compound 1), and thereby establish whether thatformulation meets the pharmacokinetic and pharmacodynamic criteria setout herein. For example, a randomized single-dose crossover study isperformed using a group of healthy adult human subjects. The number ofsubjects should be sufficient to provide adequate control of variationin a statistical analysis, and is typically about 10 or greater,although for certain purposes a smaller group suffices. Each subjectreceives administration at time zero of a single dose (e.g., a dosecontaining about 10 mg to about 300 mg of Compound 1). Blood samples arecollected from each subject prior to administration (e.g., 15 minutesbefore) and at several intervals after administration. In certaininstances, several samples are taken within the first hour and takenless frequently thereafter. Illustratively, blood samples are collectedat 0 (pre-dose), 0.25, 0.5, 1, 2, 3, 4, 6, 8, 12, and 16 hours afteradministration. If the same subjects are to be used for study of asecond test formulation, a period of at least 10 days should elapsebefore administration of the second formulation. Plasma is separatedfrom the blood samples by centrifugation and the separated plasma isanalyzed for the HDAC inhibitor (e.g. Compound 1) by a validated highperformance liquid chromatography/tandem weight spectrometry (e.g.LC-MS/MS, LC/APCI-MS/MS) procedure such as, for example, Ramu et al.,Journal of Chromatography B, 751 (2001) 49-59).

Any formulation giving the desired pharmacokinetic profile andpharmacodynamic effects is suitable for administration according to thepresent methods.

Methods of Dosing and Treatment Regimens

In one embodiment, the compositions that include an HDAC inhibitor (e.g.Compound 1) described herein are administered to humans with cancer inan amount sufficient to partially arrest the at least one of thesymptoms of the cancer. The amounts effective for this use depend on theseverity and course of the cancer, previous therapy, the patient'shealth status, weight, and response to the drugs, and/or the judgment ofthe treating physician.

In some embodiments, administration of the compound, compositions ortherapies as described herein includes chronic administration. Inspecific embodiments, chronic administration is utilized in certaininstances wherein the patient's condition does not improve and/or uponthe doctor's discretion. In certain embodiments, chronic administrationincludes administration for an extended period of time, including, e.g.,throughout the duration of the patient's life in order to ameliorate orotherwise control or limit the symptoms of the cancer.

In some embodiments, administration of the compounds, compositions ortherapies described herein is given continuously.

In some embodiments, the dose of drug being administered is temporarilysuspended for a certain length of time (i.e., a “drug holiday”). Thelength of the drug holiday varies between 4 days and 9 days.

In one aspect, pharmaceutical compositions that include an HDACinhibitor (e.g. Compound 1) are administered to humans with cancer incycles that include consecutive days of daily administration of an HDACinhibitor (e.g. Compound 1) followed by consecutive days of noadministration of an HDAC inhibitor (e.g. Compound 1). Such a dosingschedule of an HDAC inhibitor (e.g. Compound 1) allows for apharmacodynamic response to an HDAC inhibitor (e.g. Compound 1) to beachieved while limiting the incidence of Grade 4 thrombocytopenia. Grade4 thrombocytopenia typically includes instances when the human has aplatelet count less than 25,000 per mm². In one aspect, pharmaceuticalcompositions that include an HDAC inhibitor (e.g. Compound 1) areadministered to humans with cancer in cycles that include 5, 6, 7, 8 or9 consecutive days of daily administration of an HDAC inhibitor (e.g.Compound 1) followed by 5, 6, 7, 8 or 9 consecutive days of noadministration of an HDAC inhibitor (e.g. Compound 1).

If the human is receiving concurrent treatment with a second drug otherthan an HDAC inhibitor (e.g. Compound 1), then treatment with the seconddrug is not halted on the days that an HDAC inhibitor (e.g. Compound 1)is not administered. In one aspect, if the human is receiving concurrenttreatment with a second drug other than an HDAC inhibitor (e.g. Compound1), then treatment with the second drug is halted on the days that theHDAC inhibitor (e.g. Compound 1) is not administered.

In one aspect, immediate release formulations of an HDAC inhibitor (e.g.Compound 1) are administered to humans twice a day. In one aspect,immediate release formulations of an HDAC inhibitor (e.g. Compound 1)are administered to humans twice a day, the two immediate release dosesbeing administered about 3 hours to about 6 hours apart.

In one aspect, controlled release formulations of an HDAC inhibitor(e.g. Compound 1) are administered to humans once a day. In one aspect,controlled release formulations of an HDAC inhibitor (e.g. Compound 1)that are administered to humans once a day provide the same amount of anHDAC inhibitor (e.g. Compound 1) that would be obtained from dailydosing with two immediate release formulations of the HDAC inhibitor(e.g. Compound 1). In one aspect, controlled release formulations of anHDAC inhibitor (e.g. Compound 1) that are administered to humans once aday provide the same amount of an HDAC inhibitor (e.g. Compound 1) thatwould be obtained from daily dosing with two immediate releaseformulations of an HDAC inhibitor (e.g. Compound 1), wherein the twoimmediate release doses are administered about 3 hours to about 6 hoursapart.

Daily Doses

In certain embodiments, the amount of an HDAC inhibitor (e.g. Compound1), or a pharmaceutically acceptable salt thereof, that is administeredvaries depending upon factors including, by way of non-limiting example,the type of formulation utilized, the type of cancer and its severity,the identity (e.g., weight, age) of the human, and/or the route ofadministration. In various embodiments, the desired dose is convenientlypresented in a single dose or in divided doses administeredsimultaneously (or over a short period of time) or at appropriateintervals, for example as two, three, four or more sub-doses per day.

In some embodiments, the pharmaceutical compositions described hereinare in unit dosage forms suitable for single administration of precisedosage amounts. In unit dosage form, the formulation is divided intounit doses containing appropriate quantities of an HDAC inhibitor (e.g.Compound 1), or a pharmaceutically acceptable salt thereof. In oneembodiment, the unit dosage is in the form of a package containingdiscrete quantities of the formulation. Non-limiting examples arepackaged tablets or capsules, and powders in vials or ampoules. In oneembodiment, aqueous suspension compositions are packaged in single-dosenon-reclosable containers. Alternatively, multiple-dose reclosablecontainers are used, in which case it is typical to include apreservative in the composition.

Daily amounts of Compound 1, or a pharmaceutically acceptable saltthereof, which are administered to humans range from about 10 mg/mm² toabout 200 mg/mm². In one aspect, daily amounts of Compound 1, or apharmaceutically acceptable salt thereof, which are administered tohumans range from about 30 mg/mm² to about 90 mg/mm². In one aspect,daily amounts of Compound 1, or a pharmaceutically acceptable saltthereof, which are administered to humans range include about 20 mg/mm²,about 30 mg/mm², about 40 mg/mm², about 50 mg/mm², about 60 mg/mm²,about 70 mg/mm², about 80 mg/mm², or about 90 mg/mm².

In one aspect, Compound 1, or a pharmaceutically acceptable saltthereof, is administered as an immediate release formulation thatincludes about 20 mg/mm², about 30 mg/mm², about 40 mg/mm², about 50mg/mm², about 60 mg/mm², about 70 mg/mm², about 80 mg/mm², or about 90mg/mm² of Compound 1. In one aspect, Compound 1, or a pharmaceuticallyacceptable salt thereof, is administered as an immediate releaseformulation that includes about 30 mg/mm² of Compound 1, or apharmaceutically acceptable salt thereof.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered astwo immediate release formulations, where the second immediate releaseformulation is administered about 4 hours to about 6 hours after thefirst dose is administered. Each immediate release formulation includesthe same amount of the HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, as described herein. The twoimmediate release formulations provide sustained effective plasma levelsof an HDAC inhibitor (e.g. Compound 1) that are needed for therapeuticand pharmacodynamic effect while minimizing side effects. In one aspect,sustained effective plasma levels of an HDAC inhibitor (e.g. Compound 1)are maintained for about 6 hours to about 8 hours.

In one aspect, Compound 1, or a pharmaceutically acceptable saltthereof, is administered as a controlled release formulation thatincludes about 30 mg/mm², about 40 mg/mm², about 50 mg/mm², about 60mg/mm², about 70 mg/mm², about 80 mg/mm², or about 90 mg/mm² ofCompound 1. In one aspect, Compound 1, or a pharmaceutically acceptablesalt thereof, is administered as a controlled release formulation thatincludes about 60 mg/mm² of Compound 1, or a pharmaceutically acceptablesalt thereof.

In one aspect, immediate release formulations include about 10 mg toabout 300 mg of Compound 1. In one aspect, immediate releaseformulations include about 20 mg to about 200 mg of Compound 1.

In one aspect, controlled release formulations include about 20 mg toabout 600 mg of Compound I. In one aspect, immediate releaseformulations include about 40 mg to about 400 mg of Compound 1.

Cancers

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofcancer in a human. In one aspect, an HDAC inhibitor (e.g. Compound 1),or a pharmaceutically acceptable salt thereof, is used in the treatmentof a hematological cancer in a human. In one aspect, an HDAC inhibitor(e.g. Compound 1), or a pharmaceutically acceptable salt thereof, isused in the treatment of a solid tumor in a human.

Hematological cancers include cancers of the blood or bone marrow, suchas leukemia or lymphoma.

A lymphoma is a cancer that begins in cells of the immune system. Thereare two basic categories of lymphomas. One kind is Hodgkin lymphoma,which is marked by the presence of a type of cell called theReed-Sternberg cell. The other category is non-Hodgkin lymphomas, whichincludes a large, diverse group of cancers of immune system cells.Non-Hodgkin lymphomas can be further divided into cancers that have anindolent (slow-growing) course and those that have an aggressive(fast-growing) course.

A leukemia is a cancer that starts in blood-forming tissue such as thebone marrow and causes large numbers of blood cells to be produced andenter the bloodstream.

In one aspect, the cancer is a solid tumor or a lymphoma or leukemia. Inone aspect, the cancer is a carcinoma, a sarcoma, a lymphoma, aleukemia, a germ cell tumor, a blastic tumor or blastoma.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment of acancer selected from: Cardiac: sarcoma (angiosarcoma, fibrosarcoma,rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma andteratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiatedsmall cell, undifferentiated large cell, adenocarcinoma), alveolar(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus(squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma),stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductaladenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors,vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors,Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma,fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma,hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma,Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra(squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma),prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma,embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma,interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors,lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,hepatoblastom, angiosarcoma, hepatocellular adenoma, hemangioma; Bone:osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor,chordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma [pinealoma], glioblastoma multiforme,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord (neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma [serouscystadenocarcinoma, mucinous cystadenocarcinoma, endometrioid tumors,celioblastoma, clear cell carcinoma, unclassified carcinoma],granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma,malignant teratoma), vulva (squamous cell carcinoma, intraepithelialcarcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cellcarcinoma, squamous cell carcinoma, botryoid sarcoma [embryonalrhabdomyosarcoma], fallopian tubes (carcinoma); Hematologic: blood(myeloid leukemia [acute and chronic], acute lymphoblastic leukemia,chronic lymphocytic leukemia, myeloproliferative diseases, multiplemyeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin'slymphoma [malignant lymphoma]; Skin: malignant melanoma, basal cellcarcinoma, squamous cell carcinoma, Karposi's sarcoma, moles, dysplasticnevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; Adrenalglands: neuroblastoma; gallbladder carcinomas.

In one aspect, the cancer is breast cancer, colon cancer, colorectalcarcinomas, non-small cell lung cancer, small-cell lung cancer, livercancer, ovarian cancer, prostate cancer, uterine cervix cancer, urinarybladder cancer, gastric carcinomas, gastrointestinal stromal tumors,pancreatic cancer, germ cell tumors, mast cell tumors, neuroblastoma,mastocytosis, testicular cancers, glioblastomas, astrocytomas,lymphomas, melanoma, myelomas, acute myelocytic leukemia (AML), acutelymphocytic leukemia (ALL), myelodysplastic syndrome, and chronicmyelogenous leukemia (CML).

In one aspect, the cancer is a lymphoma. In one aspect, the lymphoma isa B cell lymphoma, T cell lymphoma, Hodgkin's lymphoma, or non-Hodgkin'slymphoma.

In one aspect, the cancer is a T-cell lymphoma or leukemia.

In one aspect, the T-cell lymphoma is peripheral T cell lymphoma. Inanother aspect, the T-cell lymphoma or leukemia is T cell lymphoblasticleukemia/lymphoma. In yet another aspect, the T-cell lymphoma iscutaneous T cell lymphoma. In another aspect, the T-cell lymphoma isadult T cell lymphoma. In one aspect, the T-cell lymphoma is peripheralT cell lymphoma, lymphoblastic lymphoma, cutaneous T cell lymphoma,NK/T-cell lymphoma, or adult T cell leukemia/lymphoma.

In one embodiment, the cancer is a sarcoma. A sarcoma is a cancer thatbegins in the muscle, fat, fibrous tissue, blood vessels, or othersupporting tissue of the body. Sarcomas include any one of thefollowing: alveolar soft part sarcoma, angiosarcoma,dermatofibrosarcoma, desmoid tumor, desmoplastic small round cell tumor,extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma,hemangiopericytoma, hemangiosarcoma, kaposi's sarcoma, leiomyosarcoma,liposarcoma, lymphangiosarcoma, malignant fibrous histiocytoma,neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, askin's tumor,ewing's, malignant hemangioendothelioma, malignant schwannoma,osteosarcoma, chondrosarcoma.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment of asoft tissue sarcoma in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofmyelodysplastic syndrome (MDS) in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofchronic myelogenous leukemia (CML) in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofnon-Hodgkin lymphoma in a human. In one aspect, an HDAC inhibitor (e.g.Compound 1), or a pharmaceutically acceptable salt thereof, is used inthe treatment of Hodgkin Disease in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofmultiple myeloma in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment ofchronic lymphocytic leukemia. In one aspect, an HDAC inhibitor (e.g.Compound 1), or a pharmaceutically acceptable salt thereof, is used inthe treatment of acute lymphocytic leukemia.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment of asolid tumor in a human.

In one aspect, an HDAC inhibitor (e.g. Compound 1), or apharmaceutically acceptable salt thereof, is used in the treatment of asarcoma in a human.

Combination Therapies

In one embodiment, the compositions and methods described herein arealso used in conjunction with other therapeutic reagents that areselected for their particular usefulness against the cancer that isbeing treated. In general, the compositions described herein and, inembodiments where combinational therapy is employed, other agents do nothave to be administered in the same pharmaceutical composition, and are,because of different physical and chemical characteristics, administeredby different routes. In one embodiment, the initial administration ismade according to established protocols, and then, based upon theobserved effects, the dosage, modes of administration and times ofadministration, further modified.

In certain embodiments, the particular choice of compounds used dependson the diagnosis of the attending physicians and their judgment of thecondition of the patient and the appropriate treatment protocol. Invarious embodiments, the compounds are administered concurrently (e.g.,simultaneously, essentially simultaneously or within the same treatmentprotocol) or sequentially, depending upon the nature of the cancer, thecondition of the patient, and the actual choice of compounds used. Incertain embodiments, the determination of the order of administration,and the number of repetitions of administration of each therapeuticagent during a treatment protocol, is based upon evaluation of thedisease being treated and the condition of the patient.

In one embodiment, it is understood that the dosage regimen to treat thecancer is modified in accordance with a variety of factors. Thesefactors include the type of cancer from which the human suffers, as wellas the age, weight, sex, diet, and medical condition of the human. Thus,in one embodiment, the dosage regimen actually employed varies widelyand therefore deviates from the dosage regimens set forth herein. Incertain embodiments, treatment of a cancer with a combination of an HDACinhibitor (e.g. Compound 1) and a second agent allows for the effectiveamount of the HDAC inhibitor (e.g. Compound 1) and/or the second agentto be decreased.

The formulations described herein are administered and dosed inaccordance with good medical practice, taking into account the clinicalcondition of the individual patient, the method of administration,scheduling of administration, and other factors known to medicalpractitioners.

Contemplated pharmaceutical compositions provide a therapeuticallyeffective amount of an HDAC inhibitor (e.g. Compound 1) enabling, forexample, once-a-day, twice-a-day, three times a day, etc.administration. In one aspect, pharmaceutical compositions provide aneffective amount of an HDAC inhibitor (e.g. Compound 1) enablingonce-a-day dosing.

In certain instances, it is appropriate to administer an HDAC inhibitor(e.g. Compound 1) in combination with another therapeutic agent.

In certain embodiments, the therapeutic effectiveness of an HDACinhibitor (e.g. Compound 1) is enhanced by administration of an adjuvant(i.e., by itself the adjuvant has minimal therapeutic benefit, but incombination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). In some embodiments, the benefitexperienced by a patient is increased by administering an HDAC inhibitor(e.g. Compound 1) with another therapeutic agent (which also includes atherapeutic regimen) that also has therapeutic benefit. In specificembodiments, in a treatment for cancer involving administration of anHDAC inhibitor (e.g. Compound 1), increased therapeutic benefit resultsby also providing the patient with other therapeutic agents or therapiesfor cancer. In various embodiments, administration to an individual ofan HDAC inhibitor (e.g. Compound 1) in combination with a second agentprovides the individual with, e.g., an additive or synergistic benefit.

Therapeutically-effective dosages vary when the drugs are used intreatment combinations. Determination of therapeutically-effectivedosages of drugs and other agents when used in combination treatmentregimens is achieved in any manner. For example, the use of metronomicdosing, i.e., providing more frequent, lower doses in order to minimizetoxic side effects can be utilized. In certain instances, thecombination therapy allows for either or both of the an HDAC inhibitor(e.g. Compound 1) and the second agent to have a therapeuticallyeffective amount that is lower than would be obtained when administeringeither agent alone.

A combination treatment regimen encompasses, by way of non-limitingexample, treatment regimens in which administration of an HDAC inhibitor(e.g. Compound 1) is initiated prior to, during, or after treatment witha second agent, and continues until any time during treatment with thesecond agent or after termination of treatment with the second agent. Italso includes treatments in which an HDAC inhibitor (e.g. Compound 1)and the second agent being used in combination are administeredsimultaneously or at different times and/or at decreasing or increasingintervals during the treatment period. Combination treatment furtherincludes periodic treatments that start and stop at various times toassist with the clinical management of the patient.

In any case, the multiple therapeutic agents (one of which is an HDACinhibitor (e.g. Compound 1)) are administered in any order, including,e.g., simultaneously. If administration is simultaneous, the multipletherapeutic agents are provided, in various embodiments, in a single,unified form, or in multiple forms (by way of example only, either as asingle pill or as two separate pills). In various embodiments, one ofthe therapeutic agents is given in multiple doses, or both are given asmultiple doses. In certain embodiments wherein administration of themultiple agents is not simultaneous, the timing between administrationof the multiple agents is of any acceptable range including, e.g., frommore than zero weeks to less than four weeks. In some embodiments, thecombination methods, compositions and formulations include an HDACinhibitor (e.g. Compound 1), a second agent and a third agent. Infurther embodiments, additional agents are also utilized.

In certain embodiments, the initial administration is via oraladministration, such as, for example, a pill, a capsule, a tablet, asolution, a suspension, and the like, or combination thereof. In certainembodiments, an HDAC inhibitor (e.g. Compound 1) is administered as soonas is practicable after the onset of a cancer is detected or suspected,and for a length of time necessary for the treatment of the cancer. Incertain embodiments, administration of the agents, formulations orcompositions described herein is for a length of time necessary for thetreatment of the cancer including, by way of non limiting example, forat least 2 weeks, at least 1 month, or more than 1 month.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with at least one additional therapeutic agentselected from among DNA-damaging agents; topoisomerase I or IIinhibitors; alkylating agents; PARP inhibitors; proteasome inhibitors;RNA/DNA antimetabolites; antimitotics; immunomodulatory agents;antiangiogenics; aromatase inhibitors; hormone-modulating agents;apoptosis inducing agents; kinase inhibitors; monoclonal antibodies;abarelix; ABT-888; aldesleukin; aldesleukin; alemtuzumab; alitretinoin;allopurinol; altretamine; amifostine anastrozole; arsenic trioxide;asparaginase; azacitidine; AZD-2281; bendamustine; bevacizumab;bexarotene; bleomycin; bortezomib; BSI-201; busulfan; busulfan;calusterone; capecitabine; carboplatin; carfilozib; carmustine;carmustine; celecoxib; cetuximab; chlorambucil; cisplatin; cladribine;clofarabine; cyclophosphamide; cytarabine; cytarabine liposomal;dacarbazine; dactinomycin; darbepoetin alfa; dasatinib; daunorubicinliposomal; daunorubicin; decitabine; denileukin; dexrazoxane; docetaxel;doxorubicin; doxorubicin liposomal; dromostanolone propionate;epirubicin; epoetin alfa; erlotinib; estramustine; etoposide phosphate;etoposide; exemestane; filgrastim; floxuridine; fludarabine;fluorouracil; fulvestrant; gefitinib; gemcitabine; gemtuzumabozogamicin; goserelin acetate; histrelin acetate; hydroxyurea;Ibritumomab tiuxetan; idarubicin; ifosfamide; imatinib mesylate;interferon alfa 2a; Interferon alfa-2b; irinotecan; lenalidomide;letrozole; leucovorin; leuprolide Acetate; levamisole; lomustine;meclorethamine; megestrol acetate; melphalan; mercaptopurine;methotrexate; methoxsalen; mitomycin C; mitomycin C; mitotane;mitoxantrone; nandrolone phenpropionate; nelarabine; NPI-0052;nofetumomab; oprelvekin; oxaliplatin; paclitaxel; paclitaxelprotein-bound particles; palifermin; pamidronate; panitumumab;pegademase; pegaspargase; pegfilgrastim; pemetrexed disodium;pentostatin; pipobroman; plicamycin, mithramycin; porfimer sodium;procarbazine; quinacrine; RAD001; rasburicase; rituximab; sargramostim;Sargramostim; sorafenib; streptozocin; sunitinib malate; tamoxifen;temozolomide; teniposide; testolactone; thalidomide; thioguanine;thiotepa; topotecan; toremifene; tositumomab; tositumomab/I-131tositumomab; trastuzumab; tretinoin; uracil Mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zoledronate; andzoledronic acid.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with a topoisomerase inhibitor, tubulin interactor,DNA-interactive agent, DNA-alkylating agent, and/or platinum complex.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with oxaliplatin, tyrosine kinase inhibitor,irinotecan (CPT-11), azacitidine, fludaribine, or bendamustine.

Tyrosine kinase inhibitors include, but are not limited to, erlotinib,gefitinib, lapatinib, vandetanib, neratinib, lapatinib, neratinib,axitinib, sunitinib, sorafenib, lestaurtinib, semaxanib, cediranib,imatinib, nilotinib, dasatinib, bosutinib, lestaurtinib, vatalanib andsoratinib.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with a DNA damaging anti-cancer agent and/orradiation therapy.

DNA damaging anti-cancer agents and/or radiation therapy include, but isnot limited to, ionizing radiation, radiomimetic drugs, monofunctionalalkylators (e.g. alkylsulphonates, nitrosoureas, temozolomide),bifunctional alkylators (nitrogen mustard, mitomycin C, cisplatin),antimetabolites (e.g. 5-fluorouracil, thiopurines, folate analogues),topoisomerase inhibitors (e.g. camptothecins, etoposide, doxorubicin),replication inhibitors (e.g. aphidicolin, hydroxyurea),cytotoxic/cytostatic agents, antiproliferative agents, prenyl-proteintransferase inhibitors, nitrogen mustards, nitroso ureas, angiogenesisinhibitors, inhibitors of cell proliferation and survival signalingpathway, apoptosis inducing agents, agents that interfere with cellcycle checkpoints, biphosphonates, or any combination thereof.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with an inhibitor of inherent multidrug resistance(MDR), in particular MDR associated with high levels of expression oftransporter proteins. Such MDR inhibitors include inhibitors ofp-glycoprotein (P-gp), such as LY335979, XR9576, OC144-093, R101922,VX853 and PSC833 (valspodar).

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with anti-emetic agents to treat nausea or emesis,including acute, delayed, late-phase, and anticipatory emesis, which mayresult from the use of an HDAC inhibitor (e.g. Compound 1), alone orwith radiation therapy. Anti-emetic agents include neurokinin-1 receptorantagonists, 5HT3 receptor antagonists (such as ondansetron,granisetron, tropisetron, Palonosetron, and zatisetron), GABA_(B)receptor agonists (such as baclofen), corticosteroids (such asdexamethasone, prednisone, prednisolone, or others such as disclosed inU.S. Pat. Nos. 2,789,118; 2,990,401; 3,048,581; 3,126,375; 3,929,768;3,996,359; 3,928,326 and 3,749,712), dopamine antagonists (such as,domperidone, droperidol, haloperidol, chlorpromazine, promethazine,prochlorperazine, metoclopramide), antihistamines (H1 histamine receptorantagonists, such as cyclizine, diphenhydramine, dimenhydrinate,meclizine, promethazine, hydroxyzine), cannabinoids (such as cannabis,marinol, dronabinol), and others (such as trimethobenzamide; ginger,emetrol, propofol).

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with an anti-emesis agent selected from among aneurokinin-1 receptor antagonist, a 5HT3 receptor antagonist and acorticosteroid.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with an agent useful in the treatment of anemia.Such an anemia treatment agent is, for example, a continuouseythropoiesis receptor activator (such as epoetin-α).

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered to ahuman in combination with an agent useful in the treatment ofneutropenia. Examples of agents useful in the treatment of neutropeniainclude, but are not limited to, a hematopoietic growth factor whichregulates the production and function of neutrophils such as a humangranulocyte colony stimulating factor, (G-CSF). Examples of a G-CSFinclude filgrastim.

In some embodiments, an HDAC inhibitor (e.g. Compound 1) is administeredto a human in combination with an inhibitor of at least one CYP enzyme.In situations where the HDAC inhibitor is metabolized by one or more CYPenzymes, coadministration with a CYP inhibitor reduces the in vivometabolism of the HDAC inhibitor and improves the pharmacokineticproperties of the HDAC inhibitor.

Other combination therapies are disclosed in WO 08/082,856 and WO07/109,178, both of which are herein incorporated by reference in theirentirety.

Radiation Therapy

In one aspect, an HDAC inhibitor (e.g. Compound 1) is administered incombination with radiation therapy. Radiation therapy, also calledradiotherapy, is the treatment of cancer and other diseases withionizing radiation. Ionizing radiation deposits energy that injures ordestroys cells in an area being treated (a “target tissue”) by damagingtheir genetic material, making it impossible for these cells to continueto grow. Although radiation damages both cancer cells and normal cells,the latter are better able to repair themselves and function properly.Radiotherapy can be used to treat localized solid tumors, such ascancers of the skin, tongue, larynx, brain, breast, prostate, colon,uterus and/or cervix. It can also be used to treat leukemia and lymphoma(cancers of the blood-forming cells and lymphatic system, respectively).

A technique for delivering radiation to cancer cells is to placeradioactive implants directly in a tumor or body cavity. This is calledinternal radiotherapy (brachytherapy, interstitial irradiation, andintracavitary irradiation are types of internal radiotherapy.) Usinginternal radiotherapy, the radiation dose is concentrated in a smallarea, and the patient stays in the hospital for a few days. Internalradiotherapy is frequently used for cancers of the tongue, uterus,prostate, colon, and cervix.

The term “radiotherapy” or “ionizing radiation” include all forms ofradiation, including but not limited to α, β, and γ radiation and ultraviolet light. Radiotherapy with or without concurrent or sequentialchemotherapy is an effective modality for head and neck, breast, skin,anogenital cancers, and certain nonmalignant diseases such as keloid,desmoid tumor, hemangioma, arteriovenous malformation, and histocytosisX.

Provided are methods of using an HDAC inhibitor (e.g. Compound 1) toreduce side effect caused by at least one other therapeutic treatment,such as radiation-induced normal tissue fibrosis or chemotherapy-inducedtissue necrosis, and the methods provided herein also synergisticallyinhibit tumor cell growth with radiotherapy and other anti-canceragents.

RAD51

DNA damage causes chromosomal instability, oncogensis, cell death, andsevere dysfunction of cells. The DNA repair system is cruciallyimportant for the survival of living cells. The two major DNA repairmechanisms involved in the repair of double stranded DNA breaks arehomologous recombination (HR) and non-homologous end-joining (NHEJ). Theeukaryotic RAD51 gene is an ortholog of Escherichia coli RecA, and thegene product RAD51 protein plays a central role in homologousrecombination.

Many therapeutic treatments, such as anti-cancer agents, exert theirtherapeutic effects through their capability of producing DNA damage tocells. If the cells, such as cancer cells, have active DNA repairmechanisms, the therapeutic effects of such treatments may becompromised and high dosages may be needed for achieving the desiredtherapeutic effects.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is used to decreasecellular DNA repair activity in a human with cancer.

In one aspect, presented are methods of treating cancer by using an HDACinhibitor (e.g. Compound 1) to decrease cellular DNA repair activity incombination therapy. Described are methods of combination therapy wherean HDAC inhibitor (e.g. Compound 1) interferes with a DNA repairingmechanism involving RAD51 or BRCAL

In one aspect are methods for treating cancers associated with a defectin non-homologous end joining of DNA, comprising: (a) administering to ahuman having a cancer associated with a defect in non-homologous endjoining of DNA, a therapeutically effective amount of an HDAC inhibitor(e.g. Compound 1); and (b) administering to the human a treatmentcapable of damaging cellular DNA.

The defect in non-homologous end joining of DNA comprises a defect in agene selected from the group consisting of: Ku70, Ku80, Ku86, Ku, PRKDC,LIG4, XRCC4, DCLRE1C, and XLF. In one aspect, the cancer is selectedfrom Burkitt's lymphoma, chronic myelogenous leukemia, and B-celllymphoma. In one aspect, the cancer is described herein.

In one aspect, an HDAC inhibitor (e.g. Compound 1) is used in thetreatment of an alternative lengthening of telomere (ATL) positivecancer in a human.

Additional combination therapies, treatment strategies, and the likethat include inhibiting RAD51 activity (e.g. an HDAC inhibitor (e.g.Compound 1)) are disclosed in US patent publication number 20080153877and WO 08/082,856 (both of which are herein incorporated by reference).

Kits/Articles of Manufacture

For use in the therapeutic methods of use described herein, kits andarticles of manufacture are also described herein. Such kits include acarrier, package, or container that is compartmentalized to receive oneor more containers such as vials, tubes, and the like, each of thecontainer(s) comprising one of the separate elements to be used in amethod described herein. Suitable containers include, for example,bottles, vials, syringes, and test tubes. In one embodiment, thecontainers are formed from a variety of materials such as glass orplastic.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging pharmaceutical productsinclude, e.g., U.S. Pat. Nos. 5,323,907, 5,052,558 and 5,033,252.Examples of pharmaceutical packaging materials include, but are notlimited to, blister packs, bottles, tubes, pumps, bags, containers,bottles, and any packaging material suitable for a selected formulationand intended mode of administration and treatment. A wide array offormulations of the compounds and compositions provided herein arecontemplated.

Such kits optionally comprise an identifying description or label orinstructions relating to its use in the methods described herein.

In one embodiment, a label is on or associated with the container. Inone embodiment, a label is on a container when letters, numbers or othercharacters forming the label are attached, molded or etched into thecontainer itself; a label is associated with a container when it ispresent within a receptacle or carrier that also holds the container,e.g., as a package insert. In one embodiment, a label is used toindicate that the contents are to be used for a specific therapeuticapplication. The label also indicates directions for use of thecontents, such as in the methods described herein.

In certain embodiments, the pharmaceutical compositions are presented ina pack or dispenser device which contains one or more unit dosage formscontaining a compound provided herein. The pack, for example, containsmetal or plastic foil, such as a blister pack. In one embodiment, thepack or dispenser device is accompanied by instructions foradministration. In one embodiment, the pack or dispenser is alsoaccompanied with a notice associated with the container in formprescribed by a governmental agency regulating the manufacture, use, orsale of pharmaceuticals, which notice is reflective of approval by theagency of the form of the drug for human or veterinary administration.Such notice, for example, is the labeling approved by the U.S. Food andDrug Administration for prescription drugs, or the approved productinsert.

EXAMPLES

These examples are provided for illustrative purposes only and not tolimit the scope of the claims provided herein.

Synthesis of Compound 1, HCl

Compound 1, HCl was prepared as outlined in Example 7 of U.S. Pat. No.7,276,612, the contents of which are incorporated herein by reference inits entirety.

Example 1 IV Solution of Compound 1

Compound 1 was formulated as an intravenous (IV) solutions for initialclinical trials in humans. The IV solution is an aqueous solutionformulation intended for infusion administration after dilution withisotonic saline. Each single use vial contains 25 mL of a 5 mg/mL (0.5%)solution of Compound 1 in isotonic saline and 50 mM lactate buffer, pH4.0-4.5. All the excipients in the clinical formulations are compendialand are commonly used in parenteral formulations. The quantitativecomposition of the formulation is given in Table 1. The recommendedstorage condition is 2-8° C.

TABLE 1 Quantitative Composition of IV Solution (5 mg/mL) Percent mg/gTypical Batch Ingredient (% w/w) (w/w) (57.5 kg) Compound 1 (anhydrous,free base) 0.5 5.0 0.288 kg Lactic acid 0.45 4.5 0.259 kg Sodiumchloride 0.665 6.65 0.382 kg Water for injection — — Q.S. to volume 1Nsodium hydroxide* and/or — — Q.S. to 1N hydrochloric acid* Q.S. to pH pH4.0-4.5 ± 0.2

Example 2 Immediate release Capsules

Immediate release capsules are formulated by mixing the HCl salt ofCompound 1, with microcrystalline cellulose, lactose, and magnesiumstearate and then adding the mixture into gelatin capsules (see Table2). The capsules are manufactured in two strengths. A 20 mg dosagestrength includes 20 mg of the HCl salt of Compound 1 in a size 4Swedish orange hard gelatin capsule. A 100 mg dosage strength includes100 mg of the HCl salt of Compound 1 in a size 2 dark green hard gelatincapsule. The capsules are packaged in 30 cc HDPE bottles and sealed withan induction seal and capped with a child resistant screw top cap. The20 mg dosage strength is packaged at 50 capsules per bottle. The 100 mgdosage strength is packaged at 30 capsules per bottle. The bottles arestored at controlled room temperature 20-25° C. (68-77° F.).

TABLE 2 Immediate Release Capsules Quality Component Standard Mg/CapsuleFunction Compound 1, Manufac- 20 mg^((a)) 100 mg^((a)) Active Phar- HClturer's maceutical Specification Ingredient Avicel PH113 NF 68 mg 76 mgDisintegrant (microcrys- talline cellulose) Lactose, NF 15.7 mg 17.6 mgDiluent Anhydrous Magnesium NF 1.3 mg 1.5 mg Lubricant Stearate^((a))The quantity of Compound 1 per capsule is adjusted for watercontent and purity.

Example 3 Multiparticulate Pulsatile Formulation with Timed Release

80 grams of sodium chloride and 24 grams of polyvinylpyrrolidone aredissolved in 1.2 kilograms of water and 400 grams of pulverized Compound1, HCl are suspended therein.

In a fluidized bed coater, 400 grams of starch/sugar seeds (30/50 mesh)are suspended in warm air and spray coated with the Compound 1, HClsuspension until the seeds are uniformly coated with the desired drugpotency.

Magnesium stearate in isopropyl alcohol is mixed with Eudragit NE30D(Rohm Pharma of Weiterstadt, Germany), in a proportion of two to 1 ofdried polymer to magnesium stearate. A sufficient amount of the polymersuspension is sprayed onto the active cores to provide a particular filmcoating thickness to achieve a particular lag time and rate of releasefor a population of pellets. The final coated pellets are dried at 50°C. for 2 hours to assure complete removal of moisture to stabilize thecore contents.

The procedure is repeated with at least one more batch using a differentcoating thickness to have a different lag time and rate of release. Inthis example, two populations are prepared, one with a 10% weight gainand one with a 30% weight gain of coating. Unit doses are prepared bymixing the two populations together in predetermined proportions andfilling capsules with the mixture.

After oral administration of a unit dose to a human, the firstpopulation of pellets does not begin to release Compound 1, HCl until aninitial lag time of about 2-3 hours has elapsed. The second populationof pellets does not begin to release Compound 1, HCl until an initiallag time of about 6-7 hours has elapsed. The mean release time (the timewhen half of the drug has been released) of each population of pelletsshould be separated from one another by at least 3-4 hours.

Fluidized bed coaters are well known in the art, however other coatingapparatus and methods well known in the art may be used instead.

Example 4 Alternative Multiparticulate Pulsatile Formulation with TimedRelease

The active cores are prepared as in Example 3. Magnesium stearate andtriacetin plasticizer are mixed with Eudragit RS 30D suspension in a dryweight ratio of 1:0.6:2. The polymer suspension is coated on the coresas in Example 3, preparing a plurality of populations, each having aparticular coating thickness to provide a particular lag time and rateof release of drug in an aqueous environment of use.

The different population of pellets are mixed and the mixture used tofill capsules as described in Example 3.

Example 5 Pulsatile Formulation—Tablets in Capsule

A pulsatile release dosage form for administration of Compound 1, HClsalt, is prepared by (1) formulating two individual compressed tablets,each having a different release profile, followed by (2) encapsulatingthe two tablets into a gelatin capsule and then closing and sealing thecapsule. The components of the two tablets are as follows.

TABLE 3 Tablet 1 (Without Coating) Amount per Component Function tabletCompound 1, HCl Active agent 20.0 mg Dicalcium phosphate dihydrateDiluent 38.5 mg Microcrystalline cellulose Diluent 38.5 mg Sodium starchglycolate Disintegrant  2.4 mg Magnesium Stearate Lubricant  0.6 mg

The tablets are prepared by wet granulation of the individual drugparticles and other core components as may be done using a fluid-bedgranulator, or are prepared by direct compression of the admixture ofcomponents. Tablet 1 is an immediate release dosage form, releasing theactive agent completely within 1-2 hours following administration.

Half of the immediate release tablets are coated with Delayed CoatingNo. 1 to provide Tablet 2. Tablet 2 delays the release of Compound 1,HCl by about 3-5 hours after administration. Half of the immediaterelease tablets are coated with Delayed Coating No. 2 to provide Tablet3. Tablet 3 delays the release of Compound 1, HCl by about 4-9 hoursafter administration. The coating is carried out using conventionalcoating techniques such as spray-coating or the like.

TABLE 4 Tablet 2 (with Coating) Component Function Weight Tablet 1“Core” containing the active agent 100.0 mg  Eudragit RS30D Delayedrelease coating material 8.0 mg Talc Coating component 6.0 mg Triethylcitrate Coating component 2.0 mg

TABLE 5 Tablet 3 (with Coating) Component Function Weight Tablet 1“Core” containing the active agent 100.0 mg Eudragit RS30D Delayedrelease coating material 12 mg Talc Coating component 7 mg Triethylcitrate Coating component 3.0 mg

Oral administration of the capsule to a patient should result in arelease profile having two pulses, with initial release of Compound 1,HCl occurring about 3-5 hours following administration, and release ofCompound 1, HCl from the second tablet occurring about 7-9 hoursfollowing administration.

Example 6 Pulsatile Formulation—Beads in Capsule or Tablet

The method of Example 5 is repeated, except that drug-containing beadsare used in place of tablets. Immediate release beads are prepared bycoating an inert support material such as lactose with the drug. Theimmediate release beads are coated with an amount of enteric coatingmaterial sufficient to provide a drug release-free period of about 3-5hours. A second fraction of beads is prepared by coating immediaterelease beads with a greater amount of enteric coating material,sufficient to provide a drug release-free period of about 7-9 hours. Thetwo groups of coated beads are encapsulated as in Example 5, orcompressed, in the presence of a cushioning agent, into a singlepulsatile release tablet.

Example 7 Sustained Release Tablet

Sustained release tablets of Compound 1, HCl are prepared by firstpreparing a sustained release excipient. The sustained release excipientis prepared by dry blending the requisite amounts of xanthan gum, locustbean gum, a pharmaceutically acceptable hydrophobic polymer and an inertdiluent in a high-speed mixer/granulator for 2 minutes. While runningchoppers/impellers, the water was added and the mixture was granulatedfor another 2 minutes. The granulation was then dried in a fluid beddryer to a loss on drying weight (“LOD”) of between 4 and 7%. Thegranulation was then milled using 20 mesh screens. The ingredients ofthe sustained release excipients are set forth in Table 6 below:

TABLE 6 Sustained Release Excipient Mixture Component % by WeightXanthan Gum 10 Locust Bean Gum 10 Carboxymethylcellulose 30 Dextrose 50Water  23* *removed during processing

Next, the sustained release excipient prepared as detailed above is dryblended with a desired amount of Compound 1, HCl, in a V-blender for 10minutes. A suitable amount of tableting lubricant Pruv® (sodium stearylfumarate, NF) for the following examples is added and the mixture isblended for another 5 minutes. This final mixture is compressed intotablets, each tablet containing 10% by weight, of Compound 1, HCl. Thetablets produced weighed 500 mg (Diameter is ⅜ inches; hardness is 2.6Kp). The proportions of the tablets are set forth in Table 7 below.

TABLE 7 Sustained Release Tablets Component % by Weight sustainedrelease excipient mixture of Table 6 88.5 Compound 1, HCl 10 SodiumStearyl Fumarate 1.5

Dissolution tests are then carried out on the tablets. The dissolutiontests are conducted in an automated USP dissolution apparatus (PaddleType II, pH 7.5 buffer, 50 rpm in 500 mL.). The tablets should releaseabout 30% of Compound 1, HCl by 2 hours, followed by a sustained releasesuch that about 98% of Compound 1, HCl is released at the end of 12hours.

Example 8 Coated Sustained Release Tablet

A sustained release excipient was prepared as described above by dryblending the requisite amounts of xanthan gum, locust bean gum and aninert diluent. An extra 2 minutes of granulation was used after theaddition of the components (for 4 total minutes of post-additiongranulation). Ethylcellulose aqueous dispersion was substituted forwater in the above methods. The components of the sustained releaseexcipient is described in Table 8.

TABLE 8 Sustained Release Excipient Component % by Weight Xanthan Gum 12Locust Bean Gum 18 Dextrose 65 Ethylcellulose Aqueous Dispersion  5**Ethylcellulose Aqueous Dispersion contains approx. 25% by weight ofsolids. The amount added to the formulation (i.e. 5%) is solids only.

The xanthan gum and locust bean gum are dry blended in a V-blender for10 minutes, the dextrose is added and the mixture blended for another 5minutes. The ethylcellulose aqueous dispersion is then added, followedby an additional 5 minutes of blending. The resulting granulation isthen compressed into tablets with sodium stearyl fumarate, as atableting lubricant. The tablets are then coated with additionalethylcellulose aqueous dispersion. To accomplish this, ethylcellulose(Surelease®, 400 g) is mixed with water (100 g) to form an aqueoussuspension. Thereafter, the tablets are coated in a Keith Machinerycoating pan (diameter 350 mm; pan speed 20 rpm; spray-gun nozzle 0.8 mm;tablets bed temperature 40°-50° C.; charge per batch 1 kg; dryair—Conair Prostyle 1250, 60°-70° C.). The tablets are coated to aweight gain of about 5%. The tablets should weigh about 500 mg. Theproportions of the tablets are set forth in Table 9 below:

TABLE 9 Coated Sustained Release Tablets Component % by Weight sustainedrelease excipient mixture of Table 8 83.5 Compound 1, HCl 10Ethylcellulose 5 Sodium Stearyl Fumarate 1.5

The dissolution tests are conducted in an automated USP dissolutionapparatus in such a way as to model passage through the gastrointestinaltract. The coated tablets should not release more than 10% Compound 1,HCl during the first 1-2 hours, and then should release Compound 1, HClat a steady rate such that about 90% to 100% of Compound 1, HCl isreleased after 12 hours.

Example 9 In Vitro Release Profiles

The dissolution profiles are obtained using the United StatesPharmacopeia Apparatus I at 37° C. and 100 RPM. The dissolution media isvaried with time beginning with 0.1N HCl for 0-2 hours. From 2 to 4hours the media is pH 6.5 phosphate buffer and from 4 to 24 hours themedia was PH 7.5 phosphate buffer.

Alternatively, dissolution profiles are performed using a USP Type III(VanKel Bio-Dis II) apparatus.

Example 10 In vitro Fed/Fast Dissolution Protocol

The test formulations are evaluated under a variety of dissolutionconditions to determine the effects of pH, media, agitation andapparatus. Dissolution tests are performed using a USP Type III (VanKelBio-Dis II) apparatus. In order to determine the differences, if any, indissolution kinetics between a fed state and a fasting state for theseries of formulations, in vitro dissolution experiments are carried outin a solution containing 30% peanut oil (“fed”) to model agastrointestinal tract with a typical dietary fat load. The controldetermined the dissolution rates in a solution lacking the fat load(“fasted”). The pH-time protocol (ranging from acid to alkaline to modeldigestive processes) is set forth below in Table 10, below. Agitation is15 cpm. Volume of the sample tested is 250 mL.

TABLE 10 Fed/Fast Dissolution Protocol Apparatus Media “Fed” “Fasted”Time pH 30% Peanut Oil No Peanut oil 0-1 hour 1.5 30% Peanut Oil NoPeanut oil 1-2 hour 3.5 30% Peanut Oil No Peanut oil 2-4 hour 5.5 30%Peanut Oil No Peanut oil 4-12 hour  7.5

An enteric coating on the tablet is expected to provide a tablet thatprovides dissolution rates that are not significantly different in thefasted and fed states.

Example 11 In Vitro Dose Scheduling Studies

Pharmacokinetic data (pk) data from human patients was used to modelvarious dose regimens using human cell lines: Jurkat (leukemia) andHCT-116 (colon tumor). The human cell lines were treated with or withouta HDAC inhibitor (e.g. Compound 1). Cells were cultured, treatedaccording to different regimens and concentrations to model thecorresponding dosing regimen (continuous low dose: 0.2 μM; intravenous(IV): 2 μM 3 hours+0.3 μM 4 hours; oral BID: 0.4 μM 4 hours×2, 4 hoursapart; oral consecutive BID: 0.4 μM 8 hours; oral consecutive TID: 0.266μM 12 hours), then washed out to mimic the human in vivo PK.

FIGS. 1 and 2 summarize the results of the in vitro dose schedulingstudies. It was determined that continuous exposure of the HDACinhibitor for at least 8 hours provided good efficacy. Oral bid dosingis more efficacious when given consecutively (i.e. 4 hours apart) thanwith a break in between, similar to IV dosing modeled with a 8 hexposure. Oral three times a day dosing (tid) dosing was better thantwice-a-day (bid) dosing, approaching the continuous 0.2 uM level.

Example 12 Phase 1 Trial of the Safety and Tolerability of Oral CapsuleForm of Compound 1, HCl in Advanced Cancer Patients

This is a Phase 1 dose-escalation study of the safety, pharmacokinetics,and pharmacodynamics of Compound 1, HCl administered orally in patientswith advanced cancer.

Purpose

This study seeks to determine the highest dose of Compound 1, HCl thatcan be taken without causing serious side effects in patients withadvanced cancer. The study will look at safety of the study drug(Compound 1, HCl) and whether the treatment schedule is tolerated bypatients.

Study Design

In the Phase 1 dose escalation study, up to 7 cohorts will receiveCompound 1, HCl orally at doses starting at 30 mg/m², approximately 4-6hours apart, up to 90 mg/m², 2 or 3 times a day according to 3 differentdosing schedules within 28 day cycle (5 days of dosing followed by 2days of no dosing; 5 days of dosing followed by 9 days of no dosing; 7days of dosing followed by 7 days of no dosing) until the maximumtolerated dose is reached.

Eligibility

Patients should satisfy the following criteria: At least 18 years ofage; histologically confirmed, measurable solid tumor, non-Hodgkin'slymphoma, Hodgkin's disease, chronic lymphocytic leukemia, or multiplemyeloma that has relapsed after standard therapy or for which nostandard therapy exists; ability to swallow oral capsules withoutdifficulty; estimated life expectancy>12 weeks; ECOG performancestatus≦2; Creatinine≦1.5×institutional upper limit of normal (ULN);Total bilirubin≦1.5×institutional ULN (unless elevated from documentedGilbert's syndrome); AST and ALT≦2.5×institutional ULN (≦5×institutionalULN in the presence of liver metastases); Platelet count≧100,000/μL;ANC≧1500/μL; Hgb≧9.0 g/dL; Patients with previously treated, stable,asymptomatic brain metastases who are not on corticosteroids areeligible; Willing and able to sign a written informed consent.

Results

When delivered via an immediate release capsule formulation, therapeuticeffect of Compound 1 was achieved with 2 or 3 consecutive doses (eachdose administered 4 to 6 hours apart) on days scheduled for dosing. Thedoses were given at the same time each day. Over a daily exposure rangeof 0.63 to 2.15 μM·h, three-times-a-day (TID) consecutive dosing wasassociated with a higher average grade of thrombocytopenia when comparedwith twice a day (BID) consecutive dosing. When delivered via animmediate release capsule formulation, therapeutic effect of Compound 1was achieved with 2 doses administered 4 to 6 hours apart on daysscheduled for dosing. The two doses were given at the same time eachday, with the second dose being administered approximately 4 to 6 hoursfrom the first dose.

In solid tumor and lymphoma patients, a pharmacodynamic response toCompound 1 was achieved with limited incidence of Grade 4thrombocytopenia (platelet count<25,000 per mm²) when patients receiveddrug in cycles consisting of 7 consecutive days of oral dosing followedby 7 consecutive days without dosing. In the event that a patientexperiences a treatment-related decrease in platelets to <25,000 permm², the severity of the thrombocytopenia may be ameliorated by dosingin cycles consisting 5 consecutive days of oral dosing followed by 9consecutive days without dosing.

The following pharmacokinetic information was determined for patientsthat received a 30 mg/mm² dose of the capsule formulation of Example 2.Solid tumor cancer patients received a 30 mg/mm² dose of the capsuleformulation of Example 2. Blood samples that were collected for up to 24hours post-dosing were analyzed for pharmacokinetic evaluations. Plasmawas harvested by centrifugation and stored at approximately −70° C.until analysis. Concentrations of Compound 1 were determined in plasmausing HPLC gradient system (Hewlett Packard model 1100) was configuredwith Sciex API 3000 and a reversed phase column (Phenomenex, Luna C18,3.0 μm, 50×2 mm i.d.). The mobile phase gradient consisted of 0.2%formic acid in water (A) and 0.2% formic acid in methanol (B). The flowrate was 0.4 mL/min, and run time was 2.75 minutes. Pharmacokineticanalysis was performed using WinNonlin Professional Edition (PharsightCorporation, Version 5.2). Nominal sampling times and nominal doselevels were used. The AUC_(0-4 h) for plasma concentrations of Compound1 was 0.272±0.051 μM·h (mean±SE).

The 95% confidence range for mean AUC_(0-8 h) at a dose of 60 mg/mm2 isestimated to be 0.210 to 0.742 μM·h based on a single dosing of a 30mg/mm² immediate release capsule formulation. The dose-normalized meanAUC_(0-8 h) for a once daily controlled release oral formulation iscalculated to range from 0.0035 to 0.0124 (μM·h)/(mg/m²).

Example 13 Combination Therapy: Bortezomib and Compound 1 inNeuroblastoma In Vitro and In Vivo Models

Current treatment of neuroblastoma often fails due to chemo-resistance.The current study examined the effects of Compound 1 and the proteasomeinhibitor, bortezomib in the treatment of neuroblastoma.

Neuroblastoma cell lines and patient-derived primary neuroblastomacultures were treated with bortezomib, Compound 1 alone or a combinationof both agents for 48 hours. Cells were also treated with HDACinhibitors vorinostat, sodium butyrate, and valproic acid and viabilitywas assessed by calcein AM assays. mRNA from treated cells was evaluatedat 6 and 24 hours using U133+mRNA expression arrays and Ingenuityanalysis. Dichlorofluorescein (DCF) was used to measure reactive oxygenspecies (ROS). Cell viability assays were repeated in the presence ofN-acetylcysteine (NAC). Western blot evaluated caspase-3 and PARPcleavage. Nude mice were injected with 10⁷ SMS-KCNR cells subcutaneouslyand treated with daily doses of 0.5 mg/kg bortezomib, 12.5 mg/kgCompound 1, or a combination of the two agents. Tumors were measured andimaged twice per week.

Neuroblastoma cell lines and patient cells showed sensitivity tobortezomib and Compound 1 with IC₅₀'s for bortezomib<50 nM and IC₅₀'sfor Compound 1<200 nM. The combination of bortezomib and Compound 1 wassynergistic. Expression analysis showed upregulation of NOTCH 2 and itsligands as well as c-jun. NFk-B and MYCN were both significantlydown-regulated. DCF analysis showed formation of ROS and viabilityassays showed inhibition of caspase-mediated apoptosis in the presenceof NAC. The neuroblastoma xenograft mouse model showed a decrease intumor volume in mice treated with both bortezomib and Compound 1 whencompared to the single agent treatment groups with significant survivalbenefit.

Bortezomib and Compound 1 synergistically inhibit neuroblastoma growthboth in vitro and in vivo. This combination therapy is effective andwell tolerated in the mouse model.

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes suggested to personsskilled in the art are to be included within the spirit and purview ofdisclosure and scope of the appended claims. As will be appreciated bythose skilled in the art, the specific components listed in the aboveexamples may be replaced with other functionally equivalent components,e.g., diluents, binders, lubricants, fillers, coatings, and the like.

What is claimed is:
 1. A method of treating a hematologic cancerselected from: B-cell lymphoma, Hodgkin lymphoma, chronic lymphocyticleukemia (CLL), and multiple myeloma in an individual in need thereof,comprising administering to the individual a therapeutically-effectiveamount of Compound 1

or a pharmaceutically-acceptable salt thereof.
 2. The method of claim 1,further comprising administering bendamustine.
 3. The method of claim 1,further comprising administering doxorubicin.
 4. The method of claim 1,wherein the hematologic cancer is Hodgkin lymphoma.
 5. The method ofclaim 4, further comprising administering doxorubicin.
 6. The method ofclaim 1, wherein the hematologic cancer is CLL.
 7. The method of claim6, further comprising administering bendamustine.
 8. The method of claim1, wherein the hematologic cancer is multiple myeloma.
 9. The method ofclaim 8, further comprising administering a proteasome inhibitor. 10.The method of claim 8, further comprising administering acorticosteroid.
 11. The method of claim 10, wherein the corticosteroidis dexamethasone.
 12. The method of claim 8, further comprisingadministering doxorubicin.
 13. A method of treating a sarcoma in anindividual in need thereof, comprising administering to the individual atherapeutically-effective amount of Compound 1

or a pharmaceutically-acceptable salt thereof.
 14. The method of claim13, further comprising administering bendamustine.
 15. The method ofclaim 13, further comprising administering doxorubicin.
 16. The methodof claim 13, further comprising administering a tyrosine kinaseinhibitor.
 17. A method of treating a cancer in an individual in needthereof, comprising administering to the individual: (a) atherapeutically-effective amount of Compound 1

or a pharmaceutically-acceptable salt thereof, and (b) an anti-canceragent, selected from bendamustine, a proteasome inhibitor or acorticosteroid.
 18. The method of claim 17, wherein the anti-canceragent is bendamustine.
 19. The method of claim 18, wherein the cancer isa lymphoma or leukemia.
 20. The method of claim 18, wherein the canceris chronic lymphocytic leukemia (CLL).
 21. The method of claim 17,wherein the anti-cancer agent is a proteasome inhibitor.
 22. The methodof claim 21, wherein the cancer is multiple myeloma.
 23. The method ofclaim 17, wherein the anti-cancer agent is a corticosteroid.
 24. Themethod of claim 23, wherein the corticosteroid is dexamethasone.
 25. Themethod of claim 24, wherein the cancer is a hematologic cancer.
 26. Themethod of claim 25, wherein the hematologic cancer is multiple myeloma.